INDEX

Preface.

Introduction.

PART a

Viruses: Common Characteristics.

1. Virus: Sendai RNA virus. (SeV), (SEND)
2. Virus: Rous RNA sarcoma. (RSV)
3. Virus: Adenovirus. DNA (MAV-1, MAD-1&2, FAV-1 to 8, TAV-1 & 2)
4. Virus: Cytomegalovirus. (CMV, MCMV, HHV-5, HCMV)
5. Virus: Encephalomyocarditis. -- RNA. (EMCV, TBEV)
6. Virus: Ectromelia, the DNA poxvirus of mice. (ECTV)
7. Virus: HI (Toolan) virus. (THV)
8. Virus: Sialodacryoadenitis coronavirus. RNA (SDAV)
9. Virus: Hantavirus. RNA - (HFRS, KHF, EHF, NE)
10. Virus: Coronaviruses. RNA (RCV, SDAV, PRC, IBV, MERS-COV, HCoV-, SARS-COV)

    PART b

11. Virus: Avian leukosis. (ALV/ALV-J) retrovirus
12. Virus: Rotaviruses. RNA (EDIM, ADRV, CAL) - gastroenteritis
13. Virus: Lymphocytic choriomeningitis. RNA (LCMV)
14. Virus: Theiler's encephalomyelitis. RNA (TMEV, GD VII, SAFV, VHEV)
15. Virus: Kilham rat parvovirus. DNA (RV, KRV, RPV-1, RPV-2, MKV)
16. Virus: Newcastle disease. RNA (NDV)
17. Virus: Anellovirus. DNA (TTV, TTMV, TMDV, PRA, SAV, SealAV, ZcAV)
18. Virus: Mouse Minute Parvovirus. DNA (MVM(p), MVM(i), RV, PVR-1)
19. Virus: Polyoma virus. DNA (MPV, BKV, JCV, SV40, PyV, RPV, BFD ..)
20. Virus: Norovirus. RNA (NoV, SRSVs)

PART c

• Virus: Chickenpox, varicella zoster virus (VZV)
• Virus: Hepatitis -- Liver inflammation -- 5 viruses.
• Virus: Measles, Morbilliviruses, RNA (MV, RPV)
• Virus: Reovirus type 3. RNA (ARV, MRV, BRV, NBV, RRV, )
• Virus: Pneumonia, DNA/RNS - Influenza, Syncytial, Parainfluenza. (RSV, PIV, )

PART d

Parasite: Chlamydial intracellular bacterial virus.

Bacteria: Tularemia -- Intracellular bacterium.

- Virus : Murine retroviruses. DNA (MuLV, HIV.., SMRV, MoMLV, HTLV, BLV)

- Virus : Quailpox Virus, DNA (AVP, )

Parasite: Borrelia (Deer/Bear tick)

-Focus-: Monographs on Toxins and Enhancers.

Parasite: Chlamydial intracellular bacterial virus. INDEX
http://std.about.com/od/chlamydia/f/chlabactvir.htm (2015-07-19)

LINK 02: ..//en.wikipedia.org/../Chlamydia_infection (2015-11-10)
LINK 03: http://www.ncbi.nlm.nih.gov/../PMHT0024411/ (2015-11-?)
LINK 04: http://www.petco.com/.../Chlamydia-in-Cats.aspx (2015), Cats
LINK 05: ..www.webmd.com/sexual-conditions/../chlamydia.. (2014-08-28)
LINK 06: http://chlamydia-pneumoniae.org/ (2014)
LINK 07: ..www.ncbi.nlm.nih.gov/../PMC3631192/ (2013-04-19) HHV-6
LINK 08: ..//microbewiki.kenyon.edu/../Chlamydia_trachomatis (2011-04-29)
LINK 09: ...www.skinsight.com/../chlamydialInfections.htm (2008-12-22)
LINK 10: http://iai.asm.org/../5105.long (2007-08-27), Tryptophan
LINK 11: http://www.aafp.org/afp/2006/0415/p1411.html (2006-04-15)
LINK 12: http://www.aafp.org/../p1411.html (2006-04-15), Infection
LINK 13: http://onlinelibrary.wiley.com/.../pdf (2005-09-29), Men
LINK 14: http://www.std-gov.org/stds/chlamydia.htm (2005), photos

1994 - By this year,
Chlamydial Infections begin to be associated with heart disease and other endemic North American chronic illnesses.

A South African pathologist, using an electron microscope to examine heart tissue taken during autopsies of heart disease fatalities, consistently found chlamydial infestation in the tissues. Research conducted in Helsinki found that 70% of heart attack patients in Finland had chlamydial infections and that by treating the infection the heart disease severity lessened. As the number 1 documented cause of fatalities in the USA is heart disease, one might expect that considerable resources and interest would be shown in these findings.

More than 90 million cases of Chlamydia are reported each year globally with more than a half occurred in women. The highest rate of infections is observed among teens and young adults. About half of men and most women infected with chlamydia trachomatis do not observe any symptoms which leads to the disease being untreated and easily passed from partner to partner. Chlamydia can be cured easily with simple antibiotics (if you know you have it) otherwise serious complications can occur in the reproductive system such as PID and even infertility.

Often assumed to be illnesses of aging because of their fatal frequency in patients aged 40 years and older, chlamydial influenced circulatory and systemic problems often take 10 to 30 years to develop to fatality, can be slowed in development by lifestyle changes, and are sometimes halted by either of 2 antibiotics, tetracycline and erythromycin.

2005
Chlamydiae are obligate intracellularly growing bacteria; currently four species are recognized:
C. pneumoniae, C. trachomatis, C. psittaci and C. pecorum, of the genus Chlamydia of the family Chlamydiaceae within the order Chlamydiales.

C. trachomatis is a major cause of trachoma and sexually transmitted diseases (STDs) in humans.

Two biovariants of C. trachomatis exist within the human-specific strains, which together consist of 15 serovariants. Among the trachoma biovar, serovars A, B, Ba and C are associated with ocular infection, and serovars D-K are associated with urogenital infection. Serovars L1, L2 and L3 comprise the lymphogranuloma venereum biovar.

As an intracellular parasite, still neither classified as a bacteria nor a virus, it is most often transfered from an infected host to a new host by sexual contact or during birth from mother to infant. While much of human research into heart disease, intestinal and colon irritability, and prostate and vaginal difficulties has been directed to improvements in corrective surgical sophistication or to remedial nutritional adjustments (often suggested as being "preventive"), the true origin of most such cases is chlamydial infestation and the cell and tissue damage resulting. Animals raised in a germ free environment with any selection of human diets do NOT get heart disease. This finding has not reinforced the status quo and does not sell media stories and commercial products so ... it has been largely ignored.

Genital C. trachomatis infections are among the most common STDs worldwide; it is currently estimated that c.4 million new chlamydial infections occur each year in the USA, at an estimated annual cost of >2.4 billion dollars. Approximately 70-95 percent of women and 50 percent of men with chlamydia do not observe chlamydia symptoms at all. The symptoms can also be mild and almost unnoticeable. Another reason why symptoms are not the best way to determine the infection is that it is often confused with gonorrhea as the symptoms are very much alike. Asymptomatic nature of chlamydia makes it difficult to estimate how long a person remains infectious and this period is commonly believed to last until full recovery. It can afflict the cervix in women and the urethra and rectum in both men and women. Occasionally other parts of the body (lining of the eyelid, throat and rectum) can be affected.

The spread of the infection continues to grow at geometric rates.
The incidence of chlamydial infection in women (USA) increased dramatically between 1987 and 2003, from 79 to 467 per 100,000.1 In part, this may be attributed to increased screening and improved reporting, but the burden of the disease still is significant.

There is no easy, inexpensive, accurate way to screen a human technically for the possibility of infestation. And as varieties continue to develop resistance to antibiotics, the potential for control diminishes daily.

Chlamydia is transmitted primarily through sexual activity:

unprotected intercourse (vaginal, anal) with an infected partner;

oral sex, although a less common cause of infection as bacteria Chlamydia trachomatis targets the genital area rather than the throat. Although it is possible theoretically, the cases of infestation from mouth-to-penis and penis-to-mouth contact are rare;

vagina, cervix, anus, penis or mouth contacting infected secretions or fluids which means that contraction can occur even if the penis or tongue does not enter the vagina or anus;

bacteria can travel from the vaginal area to the anus or rectum of women while wiping with toilet paper

sharing sex toys

from mother to the newborn during vaginal childbirth through the infected birth channel

infection can be transferred on fingers from the genitals to other parts of the body (for example, chlamydia can occur in the eyes)

Chlamydia is NOT contracted through simple kissing, handshaking, any casual contacts, sharing baths, towels and cups as well as from toilet seats.

The chlamydiae undergo a unique developmental cycle.
After uptake, chlamydiae develop in a so-called elementary body (EB) and grow within an intracellular vacuole, termed an inclusion. The EB is infectious but is metabolically inactive and cannot replicate. This form differentiates upon infection into the non-infectious but metabolically active replicating reticulate body (RB). Within the first 2-6 hr after internalization, EBs begin differentiating into RBs. Over the next several hours, RBs increase in number and size, until after 18-24 hr the numbers of RBs are maximal. After 24-48 hr more RBs begin differentiating back to EBs. The infected cell ruptures at 48-72 hr after infection, and the infectious EBs are released again. An intense and chronic inflammation is elicited and maintained.

Diseases caused by C. trachomatis serovars D-K in women, men and newborns.

Women
urethritis
adnexitis/PID -- pelvic inflammatory disease
long term pelvic pain 
Mucopurulent cervicitis (an inflammation of the cervix 
    with yellowish vaginal discharge and pain during sexual intercourse) 
Cystitis (inflammation of the urinary bladder) 
ectopic pregnancy (a pregnancy outside the womb)
early miscarriage or premature birth 
blocked fallopian tubes ... 
reduced fertility or infertility
perihepatitis
reactive arthritis
Reiter's syndrome


Men
urethritis
epididymitis
prostatitis
infertility
reactive arthritis
Reiter's syndrome


Newborn
preterm delivery
conjunctivitis
pneumonia

UROGENITAL INFECTION in Women
In women, chlamydial infection of the lower genital tract occurs in the endocervix.
It can cause an odorless, mucoid vaginal discharge, typically with no external pruritus, although many women have minimal or no symptoms. An ascending infection can result in pelvic inflammatory disease (PID).

Physical findings of urogenital chlamydial infection in women include cervicitis with a yellow or cloudy mucoid discharge from the os. The cervix tends to bleed easily when rubbed with a polyester swab or scraped with a spatula. Chlamydial infection cannot be distinguished from other urogenital infections by symptoms alone. Clinical microscopy and the amine test (i.e., significant odor release on addition of potassium hydroxide to vaginal secretions) can be used to help differentiate chlamydial infection from other lower genital tract infections such as urinary tract infection, bacterial vaginosis, and trichomoniasis. In addition, chlamydial infection in the lower genital tract does not cause vaginitis; thus, if vaginal findings are present, they usually indicate a different diagnosis or a coinfection.

Some women with C. trachomatis infection develop urethritis; symptoms may consist of dysuria without frequency or urgency. A urethral discharge can be elicited by compressing the urethra during the pelvic examination. Urinalysis usually will show more than five white blood cells per high-powered field, but urethral cultures generally are negative.

Women with chlamydial infection in the lower genital tract may develop an ascending infection that causes acute salpingitis with or without endometritis, also known as PID. Symptoms tend to have a subacute onset and usually develop during menses or in the first two weeks of the menstrual cycle. Symptoms range from absent to severe abdominal pain with high fever and include dyspareunia, prolonged menses, and intramenstrual bleeding. Twenty percent of women who develop PID become infertile, 18 percent develop chronic pelvic pain, and 9 percent have a tubal pregnancy. The Centers for Disease Control and Prevention (CDC) recommends that physicians maintain a low threshold for diagnosing PID and that empiric treatment be initiated in women at risk of sexually transmitted disease (STD) who have uterine, adnexal, or cervical motion tenderness with no other identifiable cause.

Culture techniques are the preferred method for detecting C. trachomatis infection, but they have been replaced in some instances by nonculture techniques. The newest nonculture technique is the nucleic acid amplification test, of which there are several. These tests have good sensitivity (85 percent) and specificity (94 to 99.5 percent) for endocervical and urethral samples when compared with urethral cultures. In women with urogenital disease, nucleic acid amplification tests can be used with an endocervical sample or a urine specimen to diagnose chlamydia.

The CDC recommends that anyone who is tested for chlamydial infection also should be tested for gonorrhea. This recommendation was supported by a study in which 20 percent of men and 42 percent of women with gonorrhea also were found to be infected with C. trachomatis.

UROGENITAL INFECTION in Men
In men, chlamydial infection of the lower genital tract causes urethritis and, on occasion, epididymitis. Urethritis is secondary to C. trachomatis infection in approximately 15 to 55 percent of men, although the prevalence is lower among older men. Symptoms, if present, include a mild to moderate, clear to white urethral discharge. This is best observed in the morning, before the patient voids. To observe the discharge, the penis may need to be milked by applying pressure from the base of the penis to the glans.

The diagnosis of nongonococcal urethritis can be confirmed by the presence of a mucopurulent discharge from the penis, a Gram stain of the discharge with more than five white blood cells per oil-immersion field, and no intracellular gram-negative diplococci. A positive result on a leukocyte esterase test of first-void urine or a microscopic examination of first-void urine showing 10 or more white blood cells per high-powered field also confirms the diagnosis of urethritis.

For diagnosis of C. trachomatis infection in men with suspected urethritis, the nucleic acid amplification technique to detect chlamydial and gonococcal infections is best (see Urogenital Infection in Women). Empiric treatment should be considered for patients who are at high risk of being lost to follow-up.

Untreated chlamydial infection can spread to the epididymis.
Patients usually have unilateral testicular pain with scrotal erythema, tenderness, or swelling over the epididymis. Men 35 years or younger who have epididymitis are more likely to have C. trachomatis as the etiologic agent than are older men.

REITER Syndrome
A rare complication of untreated chlamydial infection is the development of Reiter syndrome, a reactive arthritis that includes the triad of urethritis (sometimes cervicitis in women), conjunctivitis, and painless mucocutaneous lesions. Reactive arthritis develops in a small percentage of individuals with chlamydial infection. Women can develop reactive arthritis, but the male-to-female ratio is 5:1.

The arthritis begins one to three weeks after the onset of chlamydial infection.
The joint involvement is asymmetric, with multiple affected joints and a predilection for the lower extremities. The mucocutaneous lesions are papulosquamous eruptions that tend to occur on the palms of the hands and the soles of the feet. The initial episode usually lasts for three to four months, but in rare cases the synovitis may last about one year.

URETHRITIS is the result of initial infection with C. trachomatis in the male and can ascend from there to the contiguous urogenital organs. The clinical presentation includes dysuria, frequency and urethral pain with no voiding. Urethral discharge may or may not be present; 50-70% of infections due to C. trachomatis in males produce no symptoms. The male urethra contains all the necessary components for antigen presentation and humoral and cellular immune response. However, urethral infection with C. trachomatis induces a weak immunological response in humans.

The PROSTATITIS syndrome is one of the most common entities encountered in urological practice.
Classification of the prostatitis syndrome is based on the clinical presentation of the patient, the presence or absence of white blood cells in the expressed prostatic secretion, and the presence or absence of bacteria in the secretion. Depending upon the duration of symptoms, prostatitis is described as either acute or, where symptoms are present for +/-3 months, chronic. ...

It is generally accepted that chlamydia may ascend to the epididymis, may adhere to human spermatozoa and may enter spermatozoa. However, in any case the ejaculate has to pass through the urethra, becoming potentially contaminated on its way. ...

EPIDIDYMITIS
C. trachomatis is reported in up to 30% of patients with epididymitis.
The symptoms involve subacute to chronic scrotal pain and epididymis oedema.
Epididymitis caused by C. trachomatis can cause significant andrological sequelae.

INFERTILITY
The in vitro exposure of spermatozoa to EBs of C. trachomatis can lead to sperm death.
In vivo analyses showed that C. trachomatis is associated with sperm pathology.
C. trachomatis IgG antibodies in the man of infertile couples was related to decreased pregnancy rates and to the presence of IgG antibodies in the woman.

REACTIVE ARTHRITIS AND REITER'S SYNDROME
Especially in the male, infection with C. trachomatis can lead to reactive arthritis or Reiter's syndrome.
Predominantly peripheral joints are affected unilaterally in >60% of infections. C. trachomatis was found in up to 25% of patients with ankylosing spondylitis.

OPHTHALMIA Neonatorum
Ophthalmia neonatorum usually occurs within five to 12 days of birth but can develop at any time up to one month of age. It may cause swelling in one or both eyes with mucopurulent drainage. Prophylaxis with silver nitrate or antimicrobial ointment, which reduces the risk of gonococcal infection in neonates, does not reduce the risk of chlamydial infection.

Testing for chlamydial infection in neonates can be by culture or nonculture techniques.
The eyelid should be everted and the sample obtained from the inner aspect of the eyelid.
Sampling the exudates is not adequate because this technique increases the risk of a false-negative test.

Ophthalmia neonatorum can be treated with erythromycin base or ethylsuccinate at a dosage of 50 mg per kg per day orally, divided into four doses per day for 14 days.2 The cure rate for both options is only 80 percent, so a second course of therapy may be necessary. Topical treatment is ineffective for ophthalmia neonatorum and should not be used even in conjunction with systemic treatment.

CHLAMYDIA Pneumonia
Symptoms of chlamydial pneumonia typically have a protracted onset and include a staccato cough, usually without wheezing or temperature elevation. Findings on chest radiograph include hyperinflation and diffuse bilateral infiltrates; peripheral eosinophilia may be present.

Testing can be performed on a sample obtained from the nasopharynx.
Nonculture techniques may be used, but they are less sensitive and specific for nasopharyngeal specimens than for ocular specimens. If tracheal aspirates or lung biopsies are being collected for pneumonia in infants one to three months of age, the samples should be tested for C. trachomatis.

Like ophthalmia neonatorium, pneumonia secondary to C. trachomatis is treated with erythromycin base or ethylsuccinate at a dosage of 50 mg per kg per day orally, divided into four doses per day for 14 days.2 As with ophthalmic infection, a second course of therapy may be necessary.

SYMPTOMS:

Chlamydia Symptoms in Women:

o unusual or abnormal vaginal discharge, sometimes yellowish and smelly
o painful and frequent urinating
o bleeding between periods or heavy periods
o painful sex or bleeding after sex
o pain in the lower abdominal sometimes with nausea and low-grade fever
o swelled skin inside the vagina or around the anus

Chlamydia Symptoms in Men:

o white/cloudy, watery discharge from the tip of the penis
o painful urinating
o testicular pain and/or swelling
o swollen skin around the anus

Depending on the localization of the infection, women, men and children may experience inflamed rectum, urethra or eyelids. The symptoms of mouth and throat infections are rare although a person can suffer a sore throat. Eyes infected with chlamydia can be itchy, swelled, cause painful sensations or produce discharge similar to conjunctivitis. Infection in the rectum results in bleeding, discharge and pain.

Molecular investigation techniques for C. trachomatis have currently (2005) been accepted as the standard. The detection of C. trachomatis in prostatic secretions for the diagnosis of prostatitis has always been limited because potential contamination occurs by the passage of the specimens through the urethra. There are many kinds of tests used to support a diagnosis; many of these lack reliability or sensitivity, or, are costly in requiring very experienced lab help.

The microimmunofluorescence (MIF) test detects species- and serovar-specific antibodies that probably react with species- and serovar-specific epitopes in chlamydial MOMP. The test is highly reliable in all populations for detecting a previous exposure to chlamydiae by the presence of IgG antibodies. IgM antibodies are usually not present in patients with genital tract infections unless they represent the first exposure of the individual.

TREATMENT Options:
Chlamydia are only metabolically active in the host cell and therefore only targeted intracellularly by antibiotics. Intracellularly accumulated antibiotics are tetracyclines, macrolides and quinolones.

First-line regimens include:

--- Azithromycin 1 g orally, single dose, or
--- Doxycycline 100 mg 2-3 times a day for 10-14 days 

Alternative regimens are:

 --- Erythromycin base 500 mg orally four times a day for 7 days, or
 --- Ofloxacin 200 mg orally twice a day for 7 days, or
 --- Roxithromycin 150 mg orally twice a day for 7 days, or
 --- Clarithromycin 250 mg orally twice a dayfor 7 days 
 --- Zithromax® (azithromycin)0 gm (4 x 250 mg) a single dose, or
 --- Zithromax® Z-pak®(azithromycin)
       500mg on day 1, followed by 1 tab (250mg) once a day for 4 more days 

Although ofloxacin is generally recommended, levofloxacin has excellent activity against C. trachomatis. The treatment can consist of a single dose or last up to 2 weeks depending on the type of chlamydia.

Bacterium: Tularemia -- Intracellular bacterium. INDEX
http://wwwnc.cdc.gov/eid/article/21/1/14-0916_article (2015-01)

LINK 02: https://en.wikipedia.org/wiki/Tularemia (2015-10-03)
LINK 03: http://www.zoologix.com/.../Tularemia.htm (2015-11)
LINK 04: http://www.mayoclinic.org/.../tularemia/... (2015-07-08)
LINK 05: ..//globalbiodefense.com/...deadly-tularemia/ (2015-04-23)
LINK 06: ..//www.balkaneu.com/tularemia-the-virus-..fyrom/ (2015-03-19)
LINK 07: http://www.azdhs.gov/...presentation-colton.pdf (2015)
LINK 08: http://www.emedicinehealth.com/../page8_em.htm (2015)
LINK 09: http://www.humanillnesses.com/...Sk-Z/Tularemia.html (2015)
LINK 10: ..//www.immunology.pitt.edu/../douglas-s-reed-phd (2014-01-07)

LINK 11: ..//www.cidrap.umn.edu/.../tularemia#_Overview... (2013-09-26)
---------------- CIDRAP - Center for Infectious Disease Research and Policy
LINK 12: https://www.aphis.usda.gov/.../Tularemia...pdf (2010-10)
---------------- APHIS - Animal and Plant Health Inspection Service (USA)
LINK 13: ..//www.youtube.com/watch?v=JcRsJslrl5Q (2009-07-26) 4 min.
LINK 14: http://lubbockonline.com/stories/...Vki-CtdHAfd (2006-06-25)
LINK 15: http://www.medical-hypotheses.com/../abstract (2007-03-06)
LINK 16: http://www.azdhs.gov/phs/...tularemia.pdf (2004-08)
LINK 17: https://www.avma.org/...s/javma_222_6_725.pdf (2003-03-15)
LINK 18: http://www.state.nj.us/agriculture/.../tularemia.html (2003)
LINK 19: http://cns.miis.edu/archive/cbw/tula.htm (2001-10)

Tularemia, caused by the Gram-negative, non-motile, pleomorphic, aerobic bacillus, coccobacillus Intracellular (it is able to live as a parasite within host cells) bacterium -- Francisella tularensis (formerly known as Pasteurella tularensis). It requires cysteine (or cystine or another sulfhydryl source) for growth (although atypical strains that lack this requirement have been identified. It has a distinctive cellular fatty-acid profile.

THREE subspecies exist: Francisella tularensis tularensis (also known as Jellison type A) and Francisella tularensis holarctica (Jellison type B), and, Francisella tularensis mediasiatica.

Francisella tularensis tularensis is found in lagomorphs (rabbits, hares and pikas) in North America and is highly virulent for humans and domestic rabbits.

Francisella tularensis holarctica (or, palaearctica) is less virulent and occurs in aquatic rodents (beavers, muskrats), and voles in North America and in hares and small rodents in Eurasia. In Scandinavia, the biovar palearctica is more prevalent and can be isolated from water, aquatic mammals, and mosquitoes.

F tularensis mediasiatica is found in the Central Asian republics of the former Soviet Union (virulence is similar to subsp holarctica; infections are extremely rare); produces acid from glycerol and thus may be confused with subsp tularensis.

F tularensis is a facultative intracellular pathogen that multiplies predominantly within macrophages. The organisms initially enter macrophages (a type of white blood cell) through phagocytosis by a novel process of engulfment within asymmetric pseudopod loops and then disrupt the phagosomal membrane to gain direct access to the cytoplasm. In the blood of infected hosts, it is taken up by and replicates within leukocytes and eventually escapes in to the plasma, where it propagates a cycle of infection, escape, and reinfection. F tularensis can invade erythrocytes during infection. This feature may contribute to relapses of tularemia after short-duration antibiotic therapy, since erythrocytes have a relatively long life span (~120 days).

Outbreaks of tularemia in sheep have been reported in Montana and Idaho and can result in substantial morbidity and mortality. Outbreaks generally occur in association with reduced body condition following severe winter weather, a decreased plane of nutrition, and heavy tick infestations. Affected sheep were reported to have high rectal temperatures, low body weight, regional lymphadenopathy, and diarrhea. There is abundant seroevidence of natural infection in cattle, although a definitive clinical syndrome has not been described. In several situations, concomitant tick paralysis was observed in infected herds; however, in at least 1 epidemic, tularemia was definitively diagnosed in 2 sick calves.

A mare and 5 foals were reported to have developed tularemia, 2 of which died from the disease. The infected horses were febrile and dyspneic, had signs of depression and incoordination, and were infested with ticks; 1 of the 2 foals that died had no signs of illness. Seroconversion in surviving horses was detected, and F tularensis was isolated from tissues collected at necropsy. Livestock may be more important as maintenance hosts of the tick vectors rather than as reservoirs of infection.

People infected by F. tularensis can develop symptoms in about 3-15 days; clinical signs usually appear after 3 to 5 days. Although this disease does not cause a high number of deaths, the illness can be incapacitating for days, weeks, and sometimes even months. A sore up to 1 inch across may appear on the skin in a majority of people and is the most common sign of tularemia. If the bite associated with infection was from an animal carrying the disease, the sore is usually on the upper part of a person's body, such as on the arm. If the infection came from an insect bite, the sore might appear on the lower part of the body, such as on the leg.

Enlarged lymph nodes are seen in a majority of victims and may be the initial or the only sign of infection. Although enlarged lymph nodes usually occur as single lesions, they may appear in groups. Enlarged lymph nodes may come and go and last for as long as three years. Complications of tularemia can include pneumonia, meningitis, osteomyelitis, kidney problems, lung abscesses, pericarditis (inflammation of the sac surrounding the heart), shock, and, rarely, death.

Sweden, Finland, and Turkey have reported the highest incidences of tularemia worldwide.
In Sweden and Finland, the most common form of the disease is ulceroglandular tularemia, which is characterized by a skin ulcer at the site of infection and adjacent swollen regional lymph nodes. A marked seasonality of tularemia has been reported in Sweden; most cases occur during late summer and early autumn. Mosquito bites are the most frequent route of transmission to humans in Sweden.

In the U.S.A., Tularemia has been reported in every state except Hawaii, but a majority of cases occur in the south, central and western states, namely Alaska, Illinois, Arkansas, Oklahoma, Tennessee, Utah, Virginia and Texas. (yet) between the years 1990 - 2000, 56 percent of all reported Tularemia cases in (the USA) were in only four states: Arkansas, Missouri, South Dakota, and Oklahoma. In the United States, people living in or visiting areas of Arkansas, Missouri and Oklahoma may be at greater risk because of the concentration of ticks in those areas.

From May to October 2000, an outbreak of tularemia in Martha's Vineyard resulted in one fatality.
For a time, Martha's Vineyard was identified as the only place in the world where documented cases of tularemia resulted from lawn mowing. However, in May 2015 a resident of Lafayette, Colorado died from aerosolized F. tularensis which was connected to lawn mowing, highlighting a new vector of risk.

In Jan 2011, researchers searching for brucellosis among feral hog populations in Texas discovered widespread tularemia infection or evidence of past infection in feral hog populations of at least 2 Texas counties, even though tularemia is not normally associated with pigs at all. Precautions were recommended for those who hunt, dress, or butcher feral hogs. Since feral hogs roam over large distances, there is concern that tularemia may spread or already be present in feral hogs over a very wide geographic area.

When inhaled as an aerosol, fewer than 50 Francisella tularensis bacteria can cause disease in humans.
Aquatic animals may develop tularemia after being immersed in contaminated water. Carnivores sometimes become infected after ingesting a contaminated carcass. Vectors for F. tularensis tularensis include ticks (including Dermacentor andersoni, D. variabilis and Amblyomma americanum) and biting flies (particularly deerflies). F. tularensis holarctica is also transmitted by mosquitoes in Russia. Rarely, the organism is spread by animal bites.

F. tularensis can survive for long periods of time in arthropod vectors and in the environment.
Individual flies may carry the organism for 2 weeks and ticks throughout their lifetimes. Viable bacteria can also be found for weeks to months in the carcasses and hides of infected animals and in fomites including grain dust, straw, water, soil and bedbugs. This organism is highly resistant to freezing; live organisms have been found after 3 years in rabbit meat stored at -15° C. Both type A and B can survive for approximately 30 days in brackish water.

F. tularensis is easily killed by disinfectants including 1% hypochlorite, 70% ethanol, glutaraldehyde and formaldehyde. It can also be inactivated by moist heat (121° C for at least 15 min) and dry heat (160-170° C for at least 1 hour). Wild-type F tularensis strains generally are susceptible to aminoglycosides (streptomycin, gentamicin, kanamycin), tetracyclines, chloramphenicol, and fluoroquinolones.

SEVEN forms of tularemia are seen in humans:
typhoidal, ulceroglandular, glandular, oculoglandular, oropharyngeal and pneumonic.
The form of the disease depends on the inoculation site.

Typhoidal tularemia usually occurs after inhalation but can also develop after skin inoculation or ingestion. The clinical signs may include fever, extreme exhaustion, vomiting and diarrhea, enlarged spleen (splenomegaly), enlarged liver (hepatomegaly), pneumonia, headache, nausea, and, weight loss. Some patients become extremely weak and develop recurring chills and drenching sweats. A nonspecific rash may be seen but lymphadenopathy is usually absent. Pneumonia is particularly common in the typhoidal form and can be severe.

Ulceroglandular tularemia usually occurs after infection through the skin or mucous membranes.
The clinical signs may include fever, chills, headache, exhaustion, and, malaise. The regional lymph nodes are typically enlarged and painful; they may suppurate and drain profusely. An inflamed papule usually develops where the initial transmission occurred; it quickly turns into a pustule then ulcerates. On the extremities, single ulcers with thin, colorless, scanty exudates are usual.

In the ulceroglandular form, the organisms proliferate locally and cause a papule to develop at the site of inoculation within 3 to 5 days after initial exposure. The papule develops as a result of a localized inflammatory response that involves fibrin, neutrophils, macrophages, and T lymphocytes. The initial inflammatory nidus becomes necrotic and degenerates over the next several days, thereby forming a tender ulcerated lesion at the site of the papule. A dark scab (which may resemble the characteristic eschar of anthrax) may occur over the area of ulceration.

Organisms spread from the site of inoculation to regional lymph nodes, where they cause necrotizing lymphadenitis surrounded by a neutrophilic and granulomatous inflammatory infiltrate. Granulomas may develop in lymph nodes as the inflammatory process progresses; these may eventually coalesce to form abscesses. Follicular hyperplasia and inflammatory cell infiltrates involving predominantly granulocytes often are noted. Affected lymph nodes may become fluctuant, rupture, and sometimes create draining sinus tracts in the skin. Organisms may disseminate via hematogenous spread to involve multiple organs, and sepsis syndrome can occur.

Glandular tularemia is characterized by fever and tender lymphadenopathy without a skin ulcer.
In both glandular and ulceroglandular tularemia, organisms enter the skin through the bite of infective arthropods, direct contact with infectious materials (such as contaminated carcasses), or percutaneous inoculation with a sharp object (such as a bone fragment from a contaminated carcass). Organisms can enter through inapparent breaks in the skin surface.

Oculoglandular tularemia results from infection of the conjunctiva; this form is characterized by eye pain, eye redness, eye swelling and discharge, an ulcer on the inside of the eyelid, sensitivity to light. In some cases, there may be chemosis, periorbital edema and multiple small nodules or ulcerations on the conjunctiva.

Oropharyngeal tularemia occurs when the ulceroglandular disease occurs only in the throat.
In this form, there is fever, throat pain, mouth ulcers, vomiting, diarrhea, inflamed tonsils, swollen lymph nodes in the neck.

Septic tularemia is a severe form of the disease that affects the whole body.
Someone with this form may go into shock and experience serious health complications.
SHOCK is a condition in which blood pressure is very low and not enough blood flows to the body's organs and tissues. Untreated, shock may result in death.

Pneumonic tularemia can occur after inhalation or by secondary hematogenous spread.
Victims develop severe, sometimes fulminant, atypical pneumonia. There may be signs of lung consolidation and, in come cases, delirium. Sometimes, the only symptoms may be a dry, unproductive cough, chest pain, and, decreased breath sounds and substernal discomfort. The pneumonic form can occur with any other form and has a high mortality rate. It develops in 10 to 15% of all cases of ulceroglandular tularemia and about 50% of cases of typhoidal tularemia.

Once in the lungs, the organisms enter pulmonary macrophages within minutes and begin replicating. The explosive replicative capacity of F tularensis appears to be an important factor in virulence associated with pulmonary infection. An intense accumulation of inflammatory cells, particularly neutrophils and macrophages, can be seen at sites of bacterial replication. The influx of neutrophils appears to play more of a destructive than protective role in the host response. The following are often of notice: Ulcerative bronchitis and bronchiolitis; Discrete nodules with acute suppurative necrosis of lung parenchyma.

Also called "rabbit fever," "hunter's disease" or "deerfly fever," "Pahvant Valley plague," "Ohara's fever," -- Tularemia is caused by the bacterium "Francisella tularensis," and occurs in the United States year-round. This infectious disease was first described in Japan in 1837. Tularemia can affect all ages. Infections occur most often in hunters, butchers, farmers, fur handlers and laboratory workers.

Tularemia is named for Tulare County, California, where the organism causing the plague-like infection in golden mantled ground squirrels was first isolated in this country. Dr. Edward Francis, (1872-1957) - a graduate of Ohio State University - traveled the country for the Public Health Service, and was particularly interested in Tularemia. Francis is considered the principal researcher on the disease.

Francisella tularensis is found worldwide in over a hundred species of wild mammals, birds and insects.
The natural hosts include cottontail and jack rabbits, hares, voles, vole rats, squirrels, muskrat, beaver and lemmings. Among domestic animals, sheep seem to be particularly susceptible to clinical disease. Tularemia has also been seen in dogs, cats, pigs and horses; cattle seem to be resistant. Infections in birds, reptiles and fish have been reported.

In humans, Tularemia may (develop from) a bite from an infected wood or deer tick, deer fly, horse fly or flea, or contact with an infected rabbit. Tularemia can spread by a jab to the skin or mucous membranes with blood, tissue, or microscopic particles from the animal's skin while handling infected animals or carcasses. It can also be contracted by eating or handling insufficiently cooked rabbit meat. Less common means of spread are: drinking contaminated water, inhaling dust from contaminated soil, or handling the contaminated paws, pelts or other parts of an infected animal. Infected cats may be able to transmit the organism in bites.

People who become infected develop a painful ulceration at the site of infection, and have swollen lymph glands. Symptoms develop over about two weeks after contracting the disease, and include flu-like aches, pains and fever. If Tularemia is contracted through inhalation of bacteria, symptoms may include sudden chills, fever, abdominal pain, diarrhea, vomiting, fatigue, weight loss and headache.

Many cases may be asymptomatic.
Signs of septicemia can be seen in sheep and other mammals; symptoms may include fever, lethargy, anorexia, stiffness, increased pulse and respiration, coughing, diarrhea and pollakiuria. Rabbits and rodents may be depressed, anorectic and ataxic, with a roughened coat and tendency to huddle. Anorexia, weight loss and vomiting have been reported in cats. Skin lesions are rarely seen in animals. Symptoms usually last 2 to 10 days in susceptible animals and may end in prostration and death. Susceptible species may be found dead without other symptoms. Mortality rates up to 15% are seen in untreated lambs.

The mortality rate is approximately 30 to 35% for untreated F. tularensis tularensis infections and 5 to 15% for F. tularensis holarctica infections. Typhoidal tularemia is the most dangerous form; if untreated, the case fatality rate is approximately 35%. In contrast, the case fatality rate for the untreated ulceroglandular form is 5%. Naturally acquired cases are rarely fatal if treated. Permanent immunity can develop after a single episode of tularemia.

Most animals with acute tularemia are in good body condition.
The most consistent lesions are miliary, grayish-white necrotic foci in the liver and sometimes the spleen, bone marrow and lymph nodes. Some of these necrotic foci may be barely visible. Enlargement of the liver, spleen and lymph nodes is also common. In rabbits, the white necrotic foci on a dark, congested liver and spleen have been compared to the Milky Way. Congestion and edema is frequent in the lungs; consolidation and fibrinous pneumonia or pleuritis may also be found. The abdominal cavity sometimes contains fibrin. In some species, the lesions can resemble tuberculosis and chronic granulomas may be found in the liver, spleen, kidneys and lungs.

Although penicillin is of no use, treatment with other antibiotics is usually successful, including streptomycin. Long term immunity follows recovery from Tularemia, but re-infection has been reported. Relapses are not common but can occur if treatment is stopped before all bacteria are eliminated.

Doctors may check for F. tularensis in a blood or sputum sample that's cultured to encourage the growth of the bacteria. Sometimes tularemia can be identified by antibodies to the bacteria in a sample of blood, but these only develop several weeks after infection. You're also likely to have a chest X-ray to look for signs of pneumonia.

The laboratorial isolation of F. tularensis requires special media such as buffered charcoal and yeast extract (BCYE). It cannot be isolated in the routine culture media because of the need for sulfhydryl group donors (such as cysteine). The microbiologist must be informed when tularemia is suspected not only to include the special media for appropriate isolation, but also to ensure that safety precautions are taken to avoid contamination of laboratory personnel. Serological tests (detection of antibodies in the serum of the patients) are available and widely used. Cross reactivity with Brucella can confuse interpretation of the results, so diagnosis should not rely only on serology. Molecular methods such as PCR are available in reference laboratories.

Tularemia can be effectively treated with antibiotics such as streptomycin or gentamicin, which are given by injection directly into a muscle or vein. Depending on the type of tularemia being treated, doctors may prescribe oral antibiotics such as doxycycline (Oracea, Vibramycin, others) instead. A 14-day course of streptomycin is usually effective treatment for tularemia. Gentamicin is also effective. Although tetracycline and chloramphenicol have also been found effective, they are associated with significant relapse rates.

The inflammasome is a complex of proteins that is a dedicated killing machine for bacteria and viruses. The inflammasome that attacks the tularemia bacterium is triggered by the sensor protein AIM2 that recognizes the tularemia DNA. Both Francisella DNA and viral DNA activate AIM2 in the cell. Francisella DNA is initially detected by a DNA sensor, cGAS, which specifically engages a gene-activating protein called IRF1 in the cell.

Importantly, IRF1 triggers production of a group of proteins, called GBPs, that literally "mobs" the invading bacterium by surrounding and shredding it. The dying bacterium releases even more DNA and subsequently activates the DNA sensor AIM2 to further fuel the immune system, ultimately conquering the infection. Researchers still do not know whether GBPs directly kill bacteria or whether the GBPs require additional "help" to destroy them.

Studies with mice demonstrated the necessary role IRF1 plays in galvanizing the immune system to battle tularemia. Mice lacking IRF1 showed much higher levels of the bacteria when infected, and 100 percent succumbed to the infection, compared with only 25 percent of the mice that have IRF1. That activation of the AIM2 inflammasome by the DNA virus cytomegalovirus does not involve IRF1, suggesting that the DNA of the invading microbes is presented to the cell in different ways.

Overactivation of AIM2 has been linked to disorders including psoriasis, an abdominal aortic aneurysm and lupus. Furthermore, reduced AIM2 activity has been linked to colorectal and prostate cancers. The transcription factor IRF1 and guanylate-binding proteins target activation of the AIM2 inflammasome by Francisella infection.

Dr. Jim Alexander, regional health services zoonosis control veterinarian in Canyon, said Tularemia can be explosive.

"If the rabbits from the infected population communicate with
other populations, it could spread quite a way. Also, coyotes can
carry (infected) ticks and move it into another area."

Ken Cearley, extension wildlife specialist, said the bacterium can persist on skins or hides for more than a month and on carcasses for up to four months, so any dead animal suspected of carrying the disease should be handled with gloves.

To prevent the spread of Tularemia, hunters should also wear latex gloves when handling animals, especially rabbits, and meat must be thoroughly cooked before heading to the dinner table. Use insecticides to avoid insect bites, and don't bathe in or drink water that is untreated.

Hunters, campers, hikers, farmers, ranchers and others working outdoors in areas inhabited by rabbits and rodents are at a greater risk than other people, and these individuals should wear appropriate clothing and use insect repellent. Home gardeners and professional landscapers would best consider wearing a face mask when excavating the soil, clearing weeds or brush, or mowing lawns.

The Texas Department of State Health Services offers the following recommendations:

• Report any large die-offs of rabbits, rodents or other animals to public health agencies.
• Don't let pets roam loose
• Don't handle unknown wild animals, alive or dead (especially rabbits)
• Use insect repellent when outdoors to prevent flea and tick bites.
• Use effective flea and tick control products on pets.
• Avoid camping near rodent or rabbit burrows.
• Eliminate food and shelter sources for rodents around homes, recreational areas and work places.

Alexander said,
"While not highly susceptible, cats can get Tularemia and are more susceptible than dogs.
Horses and cattle are fairly resistant."

A long-lasting epidemic that plagued the Eastern Mediterranean in the 14th century BC was traced back to a focus in Canaan along the Arwad-Euphrates trading route. The symptoms, mode of infection, and geographical area, identified the agent as Francisella tularensis, which is also credited for outbreaks in Canaan around 1715 BC and 1075 BC. At first, the 14th century epidemic contaminated an area stretching from Cyprus to Iraq, and from Israel to Syria, sparing Egypt and Anatolia due to quarantine and political boundaries, respectively.

Subsequently, wars spread the disease to central Anatolia, from where it was deliberately brought to Western Anatolia, in what constitutes the first known record of biological warfare. Finally, Aegean soldiers fighting in western Anatolia returned home to their islands, further spreading the epidemic.

During the 1930's, the Japanese military developed and used Germ Warfare weapons in Manchuria.
It is estimated that 200,000 Chinese were killed by these weapons, of which Tularemia was one.

During World War II both Soviet Red Army and German Wehrmacht forces suffered hundreds of thousands of casualties, many from infectious disease. One of the most fiercely fought engagements in the Eurasian theater during World War II was the siege of Stalingrad, which involved at least two German group armies and resulted in the loss of millions on both sides, dead, wounded or captured.

Tularemia was deployed against Nazi troops during the battle for Stalingrad (August 1942 to February 1943). The Tularemia infections quickly arose at the beginning of the siege (and there was) a significantly high (70%) pulmonary involvement among those infected with tularemia from both sides, suggesting man-made air-borne dissemination.

A number of environmental factors would have assisted, and may have been the primary promoters of the epidemic. The Rostov region alone already had 14,000 tularemia cases in January 1942, several months before the major Panzer assault on the city. With the large epizootic pool of F. tularensis among mice and water rats (and a severely if not completely disrupted hygiene and sanitation system), (and)

".. tularemia, a disease spread by mice suddenly emerged among our pilots.
The number of infected pilots became so high, that it was necessary to
take steps to save personal structure and aircrafts:
The mice chewed all rubber and rubber insulation."

Because of its high infectivity, tularemia was developed as a BW agent by Japan, the United States and the former Soviet Union. When inhaled as an aerosol, fewer than 50 F. tularensis bacteria can cause disease in humans. Ingestion of bacteria, presumably a cause for at least a significant percentage of civilian tularemia cases during the siege of Stalingrad, requires as many as 1 billion organisms to cause disease.

... even higher rate of pulmonary involvement in tularemia infections (95.2%) during the battle of Stalingrad caused by inhaled dust from infected straw. (Also,) the war disturbed normal agricultural activities of the population in the areas near the front. Crops remained unharvested and the grass uncut. Thus was created a large source of food for rodents, and in the fall of 1942 large numbers of mice, field mice, forest mice, shrews, and others appeared in the trenches and dugouts.

The high prevalence of tularemia among Red Army soldiers was "associated with the use of hay for bedding in entrenchments, dig-outs, and trenches" and only got worse as infected rodents followed the soldiers into their hastily built military fortifications. ... many of the tularemia infections were also linked to "eating biscuits and other baked food touched by the rodents, using water from wells, which was infected by the bodies of the rodents that died as a result of tularemia." ... mosquitoes ... could have been a significant factor in the transmission of tularemia.

In the US, practical research (the BW Program) into using rabbit fever as a biological warfare agent took place in 1954 at Pine Bluff Arsenal, Arkansas, an extension of the Camp Detrick program. Tularemia was weaponized by freeze drying bacteria-laden slurry and muting it into a flue powder for aerosol delivery. The Schu S4 strain was standardized as "Agent UL" for use in the United States M143 bursting spherical bomblet. It was a lethal biological warfare agent with an anticipated fatality rate of 40 - 60%. The rate-of-action was around three days, with a duration-of-action of one to three weeks (treated) and two to three months (untreated), with frequent relapses. UL was streptomycin resistant.

The aerobiological stability of UL was a major concern, being sensitive to sunlight, and losing virulence over time after release. When the 425 strain was standardized as "agent JT" (an incapacitant rather than lethal agent), the Schu S4 strain's symbol was changed again to SR.

Both wet and dry types of F. tularensis (identified by the codes TT and ZZ) were examined during the "Red Cloud" tests, which took place from November 1966 to February 1967 in the Tanana Valley, Alaska. The United States stockpiled the bacteria during the 1960s but destroyed its stores in the 1970s at the order of the President. Russia, too, stockpiled and produced the bacteria through the mid-1990s.

The WHO (World Health Organization) estimates that if 50 kg of Tularemia bacteria compund were dispersed over a city of 5 million persons, 250,000 would become ill and 19,000 would die.

Virus: Murine retroviruses. DNA -- (MuLV, MoMLV, HTLV, BLV,
---- HIV-1, HIV-2, FIV, SIV, SMRV, MMTV) INDEX
https://en.wikipedia.org/wiki/Murine_leukemia_virus (2015-09-28)

LINK 02: ..//en.wikipedia.org/../Xenotropic_murine_leukemia..virus
LINK 03: ..//microbewiki.kenyon.edu/../Murine_Leukemia_Virus.. (2015-10-01)
LINK 04:http://www.ibc.pitt.edu/../..Murine%20Retrovirus..pdf (2015-07-13)
LINK 05: ..//www.virology.ws/../murine-gammaretroviruses-in-prostate.. (2011-06-23 )
LINK 06: http://www.ncbi.nlm.nih.gov/pubmed/18818872 (2008-09-27)
LINK 07: http://www.gvt-journal.com/content/2/1/9 (2004-08)**
LINK 08: http://www.mds-usa.com/retrovirus.html
LINK 09: ..//image.slidesharecdn.com/neurologic...murine... (1997-03-26)
LINK 10: http://jvi.asm.org/content/71/6/4531.abstract (1987-06)
LINK 11: http://www.cdc.gov/xmrv/

Retroviruses are viruses that are found throughout the animal kingdom, including in chickens, mice, cats, sheep, goats, cattle, primates, fish and humans. The first retro viruses were identified as cell free oncogenic factors in chickens. Subsequently, many of the oncogenic retroviruses have been shown to be replication defective forms that have substituted a part of their normal viral gene complement with an oncogene sequence. Replication competent retroviruses also cause malignant disease, as well as a range of other pathogenic states, in a broad range of species. This includes what must be the most significant transmissible disease of humans in recent times, acquired immunodeficiency syndrome (AIDS), which is caused by the retroviruses Human Immunodeficiency Virus Types 1 and 2 (HIV-1, HIV-2).

... many retroviruses cause life-long infections and appear to be relatively, if not completely benign, in their normal host species. In mice there are retroviruses that are very closely related to strongly oncogenic retroviruses but which are not themselves oncogenic, or are only very weakly oncogenic. In addition, there is a whole class of retroviruses, the spumaviruses, or foamy viruses, which do not appear to be linked to any specific pathogenic state. Even the simian equivalent of HIV-1, the causative agent of AIDS, is not pathogenic in all its hosts. There is also a range of endogenous retroviral sequences that are not associated with specific pathologies. Vestigial forms of retroviruses also exist; these are represented by various classes of insertional elements and can constitute a significant proportion of animal genomes.

The retroviral virion is a spherical particle.
It is enclosed by a lipid bilayer derived from the host cell plasma membrane into which one of the retroviral gene products, the envelope protein, is inserted. The virion has considerable internal structure that is mainly comprised of the products of the viral gag gene. In addition, the virion contains two identical copies of a genomic RNA molecule, a tRNA primer for reverse transcription as well as small amounts of the products of the viral pol gene. The virion may also include a range of other host cell derived proteins ...

While the simple retroviruses have only three genes, gag, pol and env, the complex retroviruses encode a number of other proteins that are involved in regulating viral replication or the host cells response to the virus. For example, HIV-1 has six gene sequences in addition to the minimal retroviral complement of gag, pol and env. Two of these, tat and rev, encode proteins that regulate expression of the viral genome, while the other four, vpu, vif, vpr and nef, encode proteins that play multiple roles in enhancing viral replication.

It is the unique nature of the retroviral life cycle, combined with the simplicity and advantageous arrangement of the retroviral genome, which has made retroviruses so attractive as vectors for gene therapy. The principal feature of the retroviral life cycle that is of interest is the ability of the retrovirus to copy its RNA genome into a double-stranded DNA form which is then efficiently and exactly integrated into the host cell genome. The integrated form is termed the provirus and it is transcribed as a normal cellular gene to produce both mRNAs encoding the various viral proteins, and the genomic RNA that is packaged into progeny virions.

The genetic structure of the virus and the existence of the proviral form make it easy to manipulate retroviruses to make replication defective vectors for transfer of heterologous gene sequences. The proviral form, being DNA, can be readily isolated in standard plasmid cloning vectors and so made amenable to molecular manipulation. The genetic structure of the virus is such that the viral cis (sequences that are biologically active in the form of nucleic acids) and trans (protein coding sequences) functions (Fig. 1) are largely non-overlapping; indeed, as far as recombinant vectors are concerned it is possible to separate them completely, albeit at some cost in efficiency.

The generation of systems capable of producing non-replication competent virus can then be achieved by placing the cis elements on a transfer vector construct and expressing the trans functions using standard recombinant plasmid expression systems. As the genomic RNA expressed from the transfer vector construct is the only RNA molecule that carries the cis signals required for packaging into the virion, and for reverse transcription and integration, no viral genes are transferred to cells infected with the resulting virus.

The resulting provirus, lacking all viral genes, is a replicative dead end and no further viral replication is possible. The nature of the retroviral replication process, where the U3 region of both the 5' and 3' LTRs of the provirus are effectively copied from the 3' LTR of the provirus in the preceding generation, also makes possible the construction of self-inactivating (SIN) vectors. With these vectors the resulting provirus contains no active retroviral derived transcriptional promoter or enhancer elements.

Murine retroviruses use at least 6 different receptors for entry into M. dunni cells:
the ecotropic, xenotropic, polytropic, amphotropic, 10A1, and Mus dunni endogenous virus groups.

Contamination of vertebrate cell lines with animal retroviruses has been documented repeatedly before. Although such viral contaminants can be easily identified with high sensitivity by PCR, it is impossible to screen for all potential contaminants. ... The first hint for the presence of contaminating retroviruses in one of our cell lines was obtained by electron microscopy of exosome-like vesicles released from the supernatants of transfected 293T cells.

Random amplification of particle associated RNAs (PAN-PCR) from supernatant of contaminated 293T cells and sequencing of the amplicons revealed several nucleotide sequences showing highest similarity to either murine leukemia virus (MuLV) or squirrel monkey retrovirus (SMRV). Subsequent mass spectrometry analysis confirmed our findings, since we could identify several peptide sequences originating from monkey and murine retroviral proteins. Quantitative PCRs were established for both viruses to test currently cultured cell lines as well as liquid nitrogen frozen cell stocks.

Gene fragments for both viruses could be detected in a broad range of permissive cell lines from multiple species. Furthermore, experimental infections of cells negative for these viruses showed that both viruses replicate rapidly to high loads. We decided to further analyze the genomic sequence of the MuLV-like contaminant virus. Surprisingly it was neither identical to MuLV nor to the novel xenotropic MuLV related retrovirus (XMRV) but showed 99% identity to a synthetic retrovirus which was engineered in the 1980s.

... The resulting sequence of the simian retrovirus confirmed our preliminary identification as squirrel monkey retrovirus with an overall sequence identity of 98,5%. In contrast the 7.4 kbp sequence of the murine retrovirus was neither identical to one of the murine leukemia viruses nor to XMLV but showed an overall similarity score of 99% to pAMS [GenBank: AF010170], a plasmid carrying the proviral sequence of a recombinant hybrid virus. This construct was engineered in the 1980s and is composed of sequences from Moloney murine leukemia virus (MoMLV) and amphotropic mouse leukemia virus clone 4070A. In the current GenBank entry it is described as "... reference retrovirus for FDA validation of retrovirus vectors used for human gene therapy...". Neither the hybrid virus itself nor the plasmid pAMS were ever used in our laboratory. ...

In summary, there have been numerous publications about retroviral contaminations like recent reports of ecotropic murine leukemia virus in various cell lines. The most frequent retrovirus found in this context is squirrel monkey retrovirus (SMRV). One study even reported the detection of SMRV related sequences in commercial interferon preparations in 1998. Although the sequences were found only as DNA and therefore rather derived from cellular DNA carrying proviral genomes than viral particles, it clearly demonstrated the contamination of the interferon producing cell line with SMRV. Germany's Central Commission of Biosafety (ZKBS) recently reported that SMRV was detectable in 128 samples of 4279 cell cultures from different laboratories throughout the country.

Defective oncogenic retroviruses .. have been described in a number of species, but have been most extensively studied in the laboratory mouse. These are replication defective, simple retroviruses in which part of the normal viral genome has been replaced with a cDNA copy of a cellular oncogene. The viral oncogene sequence often contains mutations that make the protein it encodes act in a dominant manner. The capture of a cellular oncogene by a retrovirus is an extremely rare event, the major significance of these viruses in scientific terms is that they led to the discovery of cellular oncogenes. These viruses depend on the presence of a replication competent helper virus in order to replicate and they induce cancers with relatively short latency periods.

The existence of a latency period suggests that oncogene expression is, in itself, not enough to cause malignant disease, but that additional genetic events are required. The majority of the cancers caused by these retroviruses are found in the haematopoietic system although sarcomas are also common. They are also able to transform the phenotype of cells grown in culture, principally by causing cells to lose their contact inhibition. The type of malignant event caused by any one virus is determined by the nature of the oncogene expressed by the virus and by the nature of the enhancer sequences present in the long terminal repeat which control the tissue specific expression of the oncogene. Replication defective vectors obviously also have the same potential to capture oncogenes. ...

Non-defective, replication competent retroviruses are also associated with malignant diseases.
These viruses do not carry oncogene sequences. Although first discovered in the chicken they have been most extensively studied in the laboratory mouse. These viruses induce cancer by activating cellular oncogenes via a number of different mechanisms.

In contrast to the oncogene carrying retroviruses, these viruses are associated with much longer latency periods. This is a reflection of the relatively low probability that proviral insertion will result in activation of an oncogene, in combination with the requirement for other genetic changes before a cancer eventuates. Although proviral integration can also result in gene inactivation, inactivation of tumour suppressor genes does not appear to be a mechanism associated with any known instances of retroviral induced malignancy.

The principal routes of oncogene activation are transcriptional promotion from one of the viral LTRs, and activation of endogenous cellular promoters by the strong transcriptional enhancer elements present in the viral LTRs. In the former case the provirus must obviously integrate in the sense orientation and upstream of the relevant coding sequence. Transcription can be from either LTR, and may involve splicing from either the retroviral, or cryptic, splice donor sites to a splice acceptor within the gene sequence.

If transcription is from the 3' LTR it is usually associated with inactivating mutations in the 5' LTR. Transcriptional enhancement can occur with the provirus in either orientation and over relatively large distances. This is by far the most common mechanism of oncogene activation. Another mechanism by which proviral integration can activate cellular oncogenes is by negation of negative regulatory elements in the oncogene or its transcript. However, this is a rare phenomenon. If proviral integration is downstream of the oncogene translation initiation codon a dominant variant of the oncogene product may result.

Not all non-defective simple retroviruses are overtly oncogenic and the oncogenic, non-defective simple retroviruses show a spectrum of tissue specificity and oncogenic potential. Analysis of the oncogenic potential of different retroviruses has clearly shown that the major determinant of both the overall oncogenic potential of the virus, and the cell specificity of the type of cancer that results, is the viral long terminal repeat.

More specifically, it is the transcriptional enhancer sequences in the long terminal repeat that are the major determinant of these properties. Mechanistically, this makes perfect sense. As transcriptional enhancer elements are capable of acting at a distance they will not only control transcription from the viral LTR but will also have the potential to influence transcription from promoter sequences in adjacent chromosomal genes.

In contrast to oncogene activation, the oncogenic potential of some retroviruses maps to the env gene sequences. For example, the SU protein of the polycythemic strain of Friend virus binds to, and activates, the erythropoietin receptor resulting in massive erythroid proliferation and splenomegaly. However, p55 does not bind to the active site of the Epo receptor and the Epo receptor is not used as the receptor for virus infection. In fact, p55 is not a functional envelope for infection and a helper virus is needed to allow the virus encoding p55 to propagate itself.

In an analogous manner, the sag gene of Murine Mammary Tumour Viruses (MMTV) induces an immune response by interacting with the T-cell receptor. This does not result in leukemia but facilitates the eventual induction of malignant disease in an indirect way. As the interaction between Sag and the T-cell receptor is not via the antigen binding site itself, a large proportion of the T-cell population (up to 10%) is stimulated. This, in turn, stimulates B-cells, the initial cellular target for infecting MMTV, allowing enhanced viral replication and the subsequent infection of mammary epithelial cells, the eventual site of tumour formation. Although Sag is a major determinant of the oncogenic potential of MMTV it should be noted that in the final analysis malignancy is due to oncogene activation.

How HTLV and BLV cause cancer is not entirely clear.
Both are complex retroviruses, and in addition to the gag, pol and env genes common to all retroviruses, have two genes that encode regulatory proteins. HTLV causes adult T-cell leukemia, often after a very long latency period (two or three decades can pass between infection and emergence of malignant disease). Only a small percentage of infected individuals (about 1% for HTLV) develop cancer. Although the mechanism of disease induction is unclear it is certainly related to the clonal proliferation of infected cells in vivo. Although viral gene expression does not appear to be necessary for maintenance of the disease, evidence suggests that one of the regulatory proteins, Tax, is important in inducing the initial T cell proliferation.

In terms of replication defective retroviral vectors, the study of oncogenic retroviruses suggests that oncogene activation, via the provision of promoter or enhancer sequences, but especially the latter, will be the major risk factor for disease induction. In addition, selection of the retroviral envelope used for vector pseudotyping could also potentially play a role as could inadvertent transfer and expression of other retroviral proteins, at least for vectors developed from particular retroviruses, such as Friend virus.

Several retroviruses cause CNS disease.
Some of these, such as the murine retroviruses Cas-Br-E MLV and FMCF98 are specifically associated with CNS pathology. For other retroviruses, such as HTLV and HIV, CNS disease is not the defining pathology induced by the virus, even though for the latter a high proportion of infected individuals will develop CNS disease.

Cas-Br-E MLV infects the brain via infection of the epithelial cells of the blood-brain barrier. After these become infected they release virus directly into the CNS where it infects microglial cells, resulting in a spongiform encephalopathy. The SU (env) protein has been shown to be a major determinant of the neuropathogenesis of Cas-Br-E MLV and other neuropathogenic murine retroviruses. However, the mechanisms involved have not been elucidated although receptor activation, analogous to that caused by the SU protein of the polycythemic strain of Friend virus, has been suggested but as yet remains unproven.

HTLV causes CNS disease in only a small percentage (about 1%) of infected individuals after a latent period that can be as short as two, or as long as thirty years. The development of CNS disease is not correlated with the development of ATL. For HTLV CNS disease is characterised by a vigorous inflammatory response involving T cells that causes severe demyelination in the spinal cord. Little is known about how the virus infects the CNS and what cell types are infected, or what factors influence the induction of CNS pathology.

Most individuals infected with HIV have virus within the CNS and the route of infection is thought to be transmigration of infected macrophages across the blood-brain barrier. As well as allowing opportunistic infections within the CNS there is a specific condition, AIDS dementia complex (ADC), which is a direct result of HIV infection of the CNS. Within the CNS HIV is found in macrophages and microglia, and causes demyelination, vacuolation and gliosis. Again, the mechanism by which HIV causes CNS pathology is not well understood. The gp120 (Env) and Tat proteins have been shown to be neurotoxic in vitro and a number of the cytokines induced by HIV infection of monocytes and macrophages also have the capacity to damage neural tissue, either directly or indirectly.

All of the retroviruses that cause CNS disease would appear to do so as a consequence of their active replication. In the case of HIV there is direct evidence for this -- treatment of patients with antiretrovirals can significantly decrease the severity of CNS disease. However, aspects of CNS pathology remain unresolved, for example HIV encephalitis persists even during highly active anti-retroviral therapy. Therefore, this area of retrovirus induced pathology does not appear to be of immediate relevance to replication defective retroviral vectors. However, until the mechanisms by which some aspects of CNS pathology are induced are better understood this facet of retroviral pathogenesis cannot be entirely dismissed in terms of its relevance to the design and use of retroviral vectors.

Retroviruses causing immuno-deficiencies

Simple retroviruses that cause immune deficiencies in mice, cats and primates have been described.
Somewhat surprisingly, the pathological mechanisms in these diseases are all different.

In mice, immunodeficiency is associated with proliferation of B cells (the primary target of infection), macrophages and CD4+ T-cells, all of which are non-functional. The disease is consistent with the development of anergy after antigen driven stimulation of the immune response. Expression of a mutant GAG gene product, Pr60 Gag, which is not processed normally, is required for induction of disease. However, the pathogenetic mechanisms involved are not understood. The defect in Gag processing makes the virus replication defective and a helper virus is required for virus spread, although not for induction of disease.

In cats the simple retroviruses that induce immunodeficiency do so via expression of an altered SU (Env) protein. This protein is incapable of causing resistance to superinfection; as a consequence repeated superinfection leads directly to T cell lysis and immunodeficiency then results due to a loss of T-cell function.

The lentiviruses that have been associated with immune deficiency are FIV, SIV and HIV.
All appear to share a common pathogenetic mechanism where virus infection of, and replication in, T-cells directly causes cell death, T-cell depletion and immunodeficiency. Cell death is caused by high levels of viral replication in infected cells, although the exact mechanism is unclear. However, it is also clear that the pathogenesis of HIV-1 infection is much more complicated than this, with a complex interaction between the virus and host being played out over time.

In some non-human primates, infection with SIV is usually a chronic, but largely asymptomatic, condition.
This is thought to reflect a host/virus balance that has evolved over a long period of time. Presumably, the human AIDS epidemic reflects a recent movement of HIV into the population with a resulting imbalance between viral pathogenicity and host defences, which, after a relatively long period of infection, is resolved in favour of the pathogen.

Again, the pathogenetic mechanisms involved with these retroviruses do not have major relevance to replication defective retroviral vectors. However, the pathogenetic mechanisms involved in the murine and feline immunodeficiencies caused by simple retroviruses do reiterate the point that expression of certain retroviral gene products can induce serious pathogenetic states and that this fact may have some relevance to vector design.

Implantation of retrovirus-producing cells within a tumor has been demonstrated to eliminate malignant brain tumors effectively in animal models. ... the implantation of murine fibroblast-derived virus-producing cells may induce an immune response in patients. In this study, we prepared retroviruses carrying the herpes simplex virus thymidine kinase (HTK) gene ... and stereotactically inoculated .. the HTK-bearing retroviruses into the brain tumors of mice.

Following repetitive ganciclovir (GCV) intraperitoneal injection, effective killing of glioma cells in the mouse brain was observed. The transduction efficiency was nearly as high as that observed for the implantation of high-titer retrovirus-producing fibroblasts. 80% of brain tumor-bearing mice were completely cured by our treatment protocol using concentrated HTK-harboring retroviruses. Our results suggest that repeated inoculations of high-titer retroviruses carrying the HTK gene followed by GCV treatment may be a promising strategy for the clinical treatment of malignant gliomas.

The archetypal retroviral pathogen is the oncogenic retrovirus.
Some of these are replication defective retroviruses that carry and express an oncogene sequence-indeed it was these retroviruses that largely allowed the concept of oncogenes to be first defined. These viruses induce cancers with relatively short latency periods. In addition, there are a large number of non-defective retroviruses that are oncogenic. These generally induce cancers after longer latency periods.

HTLV and BLV and related viruses form a separate class of complex retroviruses that cause leukemia in a small percentage of infected individuals after very long latency periods. Retroviruses have also been associated with sarcomas in fish but these viruses have not been studied in great detail.

The preferential infection and integration of replication-deficient retroviral vectors into the genome of actively dividing cells facilitates their utilization in the transduction of the HTK gene into brain tumors in clinical trials of gene therapy in the treatment of malignant brain tumors. ...

A unique property of the HTK/GCV system has been discovered while treating brain tumors.
Tumor cells that did not express the HTK suicide gene were also eliminated together with the HTK-bearing cells. Hence, this effect has been designated the bystander effect because HTK-positive cells can induce the death of HTK-negative neighboring (or bystander) cells. Through the bystander effect, effective tumor ablation is achieved, if merely 10-20% of the tumor cells contain the HTK gene. ...

We initially developed new virus-producing cells to obtain a high-titer virus supernatant through the introduction of the polyoma early gene. Activation of the LTR (long terminal repeat) promoter by the polyoma early gene products increased the titer of the cell culture medium ... before concentration. ...

Kuriyama et al reported the complete eradication of hepatocellular carcinoma in subcutaneous tissues of mice by repeated injections of retroviruses. ... The direct intracranial inoculation of the reagent is limited by its volume because of the risk of increasing intracranial pressure. We have often observed that slight increases in intracranial pressure prove critical for brain tumor patients. This report, therefore, provides the first evidence for the promising applicability of repeated direct inoculations of concentrated retrovirus harboring the HTK gene followed by GCV treatment in the eradication of malignant gliomas in the brain.

Virus: Quailpox Virus, DNA (AVP, ) INDEX
https://en.wikipedia.org/wiki/Avipoxvirus (2015)

LINK 2: http://avianmedicine.net/content/uploads/2013/03/32.pdf (2013)
LINK 3: http://viralzone.expasy.org/all_by_species/151.html
LINK 4: http://ps.oxfordjournals.org/content/64/1/65.abstract (1984-02-21)
LINK 5: ..//waryofvirus.blogspot.ca/../virus-famili-poxviridae.html (2011-10)
LINK 6: http://www.cdc.gov/poxvirus/diseases.html
LINK 7:

Members of the Poxviridae family (Avipoxvirus genus) cause a variety of diseases in birds.
These large DNA viruses (250 to 300 nm) induce intracytoplasmic, lipophilic inclusion bodies called Bollinger bod- ies (pathognomonic). These inclusion bodies may be identified in affected epithelial cells of the integument, respiratory tract and oral cavity.

The Avipox genus has a high capacity for recombination, which has been shown to occur between field and vaccine strains of virus when actively infected flocks are vaccinated. Many experimental infections have been performed without determining the strain of virus, which probably adds to confusion about poxvirus epizootiology.

The effects of this virus might increase the difficulty of breathing, eating, and drinking significantly. Exterior lesions are restricted to the non-feathered parts of the body, usually the face and legs and are characterized by proliferative and necrotizing dermatitis.

Unlike other viruses, avipoxvirus can withstand extreme dryness.
With this advantage, it can spread on dust particles.
This is because it has adapted to living in the dry mucous membranes of an infected host's upper respiratory tract.

Avian poxvirus infections, particularly in a flock situation, can remain latent for years.
Non-specific stress factors are associated with viral reactivation.
It has been suggested that latent poxvirus infections (including vaccine strains) can be egg transmitted (at least in the chicken). Fowlpoxvirus (including vaccine strains) is known to induce a mild immunodepression that potentiates secondary infections.

Avipoxviruses infect a number of domestic and wild birds and can be identified as causing disease in at least 232 species in 23 orders. Transmission usually occurs by skin abrasions, inhalation, or by biting insects, such as mosquitoes.

Avipoxviruses
Canarypox virus 
Fowlpox virus 
Juncopox virus 
Mynahpox virus 
Pigeonpox virus 
Psittacinepox virus 
Quailpox virus 
Sparrowpox virus 
Starlingpox virus 
Turkeypox virus 

Tentative members
Crowpox virus 
Peacockpox virus 
Penguinpox virus 

The infection of avipoxvirus can lead to decreased egg production, reduced growth, blindness, and increased mortality in domestic poultry. In wild bird species, elevated predation secondary infections, trauma, reduced male mating success, and death are common results.

Avian pox viruses are contagious pathogens, and once introduced into a captive community, can spread very quickly.

LIFE CYCLE
Viral replication is cytoplasmic.
Entry into the host cell is achieved by attachment of the viral proteins to host glycosaminoglycans (GAGs) mediates endocytosis of the virus into the host cell. Fusion with the plasma membrane to release the core into the host cytoplasm.

Early phase: early genes are transcribed in the cytoplasm by viral RNA polymerase.

Early expression begins at 30 minutes post-infection.
Core is completely uncoated as early expression ends, viral genome is now free in the cytoplasm.

Intermediate phase:
Intermediate genes are expressed, triggering genomic DNA replication at approximately 100 minutes post-infection.

Late phase:
Late genes are expressed from 140 min to 48 hours post-infection, producing all structural proteins.
Assembly of progeny virions starts in cytoplasmic viral factories, producing an spherical immature particle.

This virus particle matures into brick-shaped intracellular mature virion (IMV).
IMV virion can be released upon cell lysis, or can acquire a second double membrane from trans-Golgi and bud as external enveloped virion (EEV) host receptors, which mediates endocytosis.

Replication follows the DNA strand displacement model.
DNA templated transcription is the method of transcription.
The virus exits the host cell by existing in occlusion bodies after cell death and remaining infectious until finding another host.

Birds serve as the natural host.
Transmission routes are mechanical, contact, and air borne particles.

Cutaneous Form ("Dry Pox"):
The cutaneous form is the most common form of disease in many raptors and Passeriformes but not in Psittaciformes. Changes are characterized by papular lesions mainly on unfeathered skin around the eyes, beak, nares and distal to the tarsometatarsus. The interdigital webs are most frequently affected in waterfowl and the Shearwater. As lesions progress, papules change color from yellowish to dark brown and develop into vesicles that open spontaneously, dry and form crusts. Spontaneous desquamation may require weeks and occurs without scarring in uncomplicated cases. Pigmented skin will frequently be discolored following an infection. Secondary bacterial or fungal colonization of lesions can substantially alter the appearance and progression of the disease.

Avian Pox: Infection and Immunity with Quail, Psittacine, Fowl, and Pigeon Pox Viruses by R. W. WINTERFIELD and W. REED --- February 21, 1984

Quail, chickens, and turkeys vaccinated with pigeon and fowl pox viruses were not protected against challenge of their immunity with quail pox virus and they developed severe cutaneous lesions of pox. When quail and chickens were vaccinated with quail pox virus and given pigeon and fowl pox challenge viruses, no protection was present.

Thus, quail pox virus had no immunologic relationship to pigeon and fowl pox viruses.

Psittacine pox virus applied as a vaccine in quail and chickens also failed to protect against quail pox virus challenge. However, quail, chickens, and turkeys vaccinated with quail pox virus were protected against quail pox virus challenge.

An isolate of psittacine pox virus, applied as a vaccine, protected chickens against challenge with the same virus isolate and also against challenge with two other psittacine pox virus isolates, confirming a close or identical antigenic relationship with each other.

When combined in a multivalent vaccine, quail, psittacine, and fowl pox viruses induced excellent protection in chickens against challenge with the three respective viruses.

Acyclovir has proven to be effective with some strains of avian herpesvirus and may have positive effects in treating poxvirus infections.

An unequivocal diagnosis of a particular viral infection can be made only through specific laboratory diagnostic methods. Clinical, pathologic and histologic changes are perhaps suggestive of a diagnosis but pathognomonic lesions are rare, and in-depth diagnostic tools are necessary to confirm a virus as a cause of morbidity or mortality in an avian host.

Parasite: Borrelia (Deer/Bear tick) INDEX
https://en.wikipedia.org/wiki/Ixodes_scapularis (2015-10-19)

LINK 02: http://www.ei-resource.org/.../lyme-disease/ (2015-10-20)
LINK 03: http://geology.com/articles/ticks-lyme-disease/ (2015)
LINK 04: http://www.cdc.gov/lyme/.../TickborneDiseases.pdf (2015)
LINK 05: ..//www.health.state.mn.us/../tickborne/ticks.html (2014-04-25)
LINK 06: http://borreliaburgdorferi.org/ (2014) What is
LINK 07: ..//microbewiki.kenyon.edu/../Borrelia_burgdorferi_and_Lyme.. (2013-05-08)
LINK 08: http://www.textbookofbacteriology.net/Lyme.html (2012) 6 pages

LINK 09: ..//www.health.state.mn.us/.../twoyrcycle.html (2011-01-14)
LINK 10: http://www.scielo.br/.../v40n4/6497.pdf (2007) Brazil **
LINK 11: https://www.ncbi.nlm.nih.gov/pubmed/15665404 (2004) Brain
LINK 12: http://www0.nih.go.jp/JJID/57/229.pdf (2004-12) East Asia
LINK 13: https://www.ncbi.nlm.nih.gov/../15893422 (2005-08-12) Pathology
LINK 14: https://www.ncbi.nlm.nih.gov/../12653133 (2001, Spring)
LINK 15: https://www.ncbi.nlm.nih.gov/../10672174 (2000-02) Genome

Lyme disease is an infectious disease caused by a spirochete (spiral shaped bacterium) known as Borrelia burgdorferi (Bb). Lyme disease is most often spread by ticks, but can also be transmitted by fleas, mosquito's, and mites. Evidence has suggested that these insects don't actually need to bite you for you to become infected. Worryingly, there is also evidence that Lyme disease can be spread by a number of other non-insect methods including from person to person through sex, or from mother to baby in the womb.

Researchers at the University of Wisconsin have found dairy cattle and other animals can acquire he disease and pass it on to humans through the food chain. The Centers for Disease Control (CDC) in Atlanta, believe that Borrelia burgdorferi can even survive the blood purification processes that donated blood is subjected to, and the disease can thus be acquired through receiving a blood transfusion.

Lyme Disease is considered to be one of the fastest growing illnesses in the world.

Lyme disease is said to be rampant in the US with an estimated 200,000 new cases every year and and estimated 1.8 million Americans infected, according to the CDC. With awareness of the disease and appropriate testing methods still low, actual infection numbers may be higher still. One Lyme disease expert, Dan Kinderleher, MD has publicly stated that the actual number of Americans infected may even be 10 times the 1.8 million reported by the CDC. Doctors in Europe are currently less likely to look for Lyme, but positive samples have been detected in European countries including England, Scotland, Ireland, France, Germany, Spain, Switzerland and Denmark.

The Symptoms of Lyme Disease

It is well known that a large proportion of individuals infected with the Borrelia burgdorferi spirochete, do not show any signs of Lyme disease at all. In a 1998 study conducted in Switzerland, it was found that only 12.5% of those infected with Bb had any clinical symptoms. All carriers of Bb are at risk of developing Lyme disease at some time however. The bacteria may lie dormant for many years but be activated by events such as increased stress or another infection which alter immune activity.

When symptoms are present, Lyme disease is a master of disguise.
Lee Cowden, MD states that there are very few symptoms or unexplained illnesses (or even already diagnosed illnesses!) where Lyme should not be considered a factor. The Sierra Integrative Medicine Clinic in Reno, Nevada, states that

"Authorities estimate that up to 90% of the (US) population could be carrying the Lyme spirochete and that Lyme is a factor in over 50% of chronic illnesses."

Dr. Atanas Tzonkov, director of Bulgaria's largest private medical clinic, has successfully treated thousands of patients for Lyme infection. He says that these patients had been diagnosed with over 100 different conditions before he treated them. A possible explanation for this is that most of these patients 2 of 15were actually suffering from misdiagnosed Lyme disease, or, that Lyme infection is actually a component of these many different illnesses. Lyme disease can obviously mimic a large number of illnesses through the production of a wide range of symptoms. Some tell-tale signs of Lyme disease however, include:

Direct tick bite related symptoms:

A red spot around the location of the tick's bite.
Spots gradually grow bigger, often with a pale area in the middle.
This is known as erythema migrans.

Erythema migrans may sometimes appear at other places on the body where you have not been bitten.
Some people develop many red spots.

Usually one to four weeks will pass between the bite and when erythema migrans appears.

Initial Systemic Lyme disease symptoms:

o - Fatigue
o - Joint and muscle pains
o - Mild fever
o - Headaches
o - Drowsiness
o - Swollen lymph glands

Lyme disease patients often describe their symptoms as "feeling like I have flu", a description commonly used by chronic fatigue syndrome and fibromyalgia patients to describe their illness.

The Canadian Lyme Disease Foundation lists a total of 75 symptoms in multiple body systems that infection with Bb may cause. Some of these include:

Head, Face & Neck

o - Unexplained hair loss
o - Headache, mild or severe, Seizures
o - Pressure in Head, White Matter Lesions in Head (MRI)
o - Twitching of facial or other muscles
o - Jaw pain or stiffness

Eyes & Vision

o - Double or blurry vision
o - Pain in eyes, or swelling around eyes
o - Light Sensitivity

Musculoskeletal System

o - Bone pain, joint pain or swelling, carpal tunnel syndrome
o - Stiffness of joints, back, neck, tennis elbow
o - Muscle pain or cramps, (Fibromyalgia)

Neurological System

o - Tremors or unexplained shaking
o - Burning or stabbing sensations in the body
o - Fatigue, Chronic Fatigue Syndrome, Weakness, peripheral neuropathy or partial paralysis
o - Pressure in the head
o - Numbness in body, tingling, pinpricks

Psychological Well-being

o - Mood swings, irritability, bi-polar disorder
o - Unusual depression
o - Disorientation (getting or feeling lost)
o - Feeling as if you are losing your mind
o - Over-emotional reactions, crying easily

Cognitive Symptoms

o - Memory loss (short or long term)
o - Confusion, difficulty in thinking
o - Difficulty with concentration or reading
o - Going to the wrong place

General Well-Being

o - Unexplained fevers (high or low grade)
o - Continual infections (sinus, throat, chest, etc.)
o - Low body temperature
o - Allergies & Chemical Sensitivities
o - Increased affect from alcohol and possible worse hangover

How Lyme Infection Causes Disease - Bio-Toxins Induced Illness

... Microbial toxins can damage the body through both directly damaging tissues and indirectly through toxemia (the presence of toxins in the blood stream). ..

The term "Bio-toxins induced illnesses" has recently been introduced in light of this research into microbial toxins, and Lyme disease is considered as the most important in this category.

On the subject of molecular toxicology, Dr. C. Shoemaker, MD and H Kenneth Hudnell, PhD state that "Borrelia burgdorferi produces a large suite of bio-toxins that have tissue affinity, mainly neurotoxins with high molecular tropism for lipid structures (i.e., central nervous system, peripheral nervous system), muscles, joints, lungs and many others." These statements are backed up by research data identifying specific neurotoxins from Bb.

What this means is that Borrelia burgdorferi toxins are strongly attracted to fatty cells (such as those of the nervous system), and exert effects that disrupt normal functioning of these cells. These bio-toxins are thought to alter various specific sites in the brain on molecular, structural and chemical levels, interfering with all the major neurotransmitters (dopamine, serotonin, norepinephrine, acetylcholine, GABA).

In this way they can cause all manner of neurological and psychiatric symptoms that mimic (or cause) many illnesses, from degenerative diseases such as Parkinson's Disease and Alzheimer's, to depression and autistic disorders. Indeed, Dr. Paul Fink, a former president of the American Psychiatric Association, has acknowledged that Lyme disease can contribute to every single psychiatric disorder in the Diagnostic Symptoms Manual IV (DSM-IV), the manual used to diagnose psychiatric disorders.

Lyme bio-toxins are also known to interfere with many hormones, enzymes and their receptors, interfering with their usual functioning. One symptom that is prevalent in Lyme disease and related illnesses (chronic fatigue syndrome, fibromyalgia, autism, depression etc), is chronic fatigue. This can be explained by molecular toxicology research that shows that the calcium channel's normal functioning may be altered by Bb's bio-toxins. This results in the impairment of enzymes and other cellular chemicals involved in cellular energy production.

The same may be true for muscle and joint tissues, resulting in the aches and pains associated with Lyme disease. In fact there is a type of arthritis associated with Borrelia burgdorferi, known as Lyme Arthritis. This condition is officially reported as being rare, but as with all Lyme associated illnesses, accurate diagnosis and reporting may be well below actual numbers.

Diagnosing Lyme Disease

Diagnosing Lyme disease can be a very difficult task, which is why so many cases are missed, or patients are diagnosed with another illness entirely. For starters, only about 30% of people infected develop the tell-tale 'bulls-eye' rash (erythema migrans) from a tick bite, and even then, physicians often miss it.

Sometimes the rash may not present as a 'bulls-eye' but will be more generalized.
Patients often don't even remember being bitten by a tick as they often, especially in the nymph phase, are able to feed and drop off unnoticed. The incubation period from infection to onset of erythema migrans, is typically 7 to 14 days, but may be as short as 3 days or as long as many years.

The Canadian Lyme Disease Foundation reports that

"Time and again this office receives phone calls from patients who have a rash, and in some cases had had a tick attached at the site, only to be told by their physician to come back a month later for a test. It is imperative that clinicians understand that the presence of a homogenous or "bull's eye" rash caused by a tick-bite is indicative of infection, and treatment should be started immediately. Do not look at this as simply an allergic reaction to the bite. Err on the side of caution. Most blood tests do not work until 4-6 weeks after infection has occurred, and any delay may cause complications at a later date"

There are a number of laboratory tests that doctors may use to try to detect Lyme disease but they are notoriously inaccurate, often producing "false negative" results, i.e. showing that a patient does not have Lyme disease when in fact they do. The reasons for this are due to the fact that the Borrelia burgdorferi spirochete is so adept at evading both our immune cells and conventional bacterial detection methods.

Like other tick-borne organisms, Borrelia burgdorferi, does not have one single, static appearance or chemical signature, but is able to alter these in order to evade detection. It is thought to have a number of genes whose purpose it is to randomly alter the organisms outer surface proteins (OSP's) so as to evade the immune system's fighters. In this way, Borrelia burgdorferi also often evades standard laboratory testing procedures.

Add to this ability, the fact that there are 300+ known strains of Bb, and it becomes obvious that this is one elusive bug. It is not uncommon for a patient to display the bulls-eye rash of erythema migrans, and all the other core symptoms of recent Bb infection, only to repeatedly produce negative lab test results.

The Laboratory Tests:

EIA, ELISA, IFA or PCR-DNA probe
- According to Dr. Charles. L. Crist, an experienced Lyme disease expert, one of these tests will be the first line of testing used by the average doctor who suspects Lyme. Unfortunately, these tests are not very accurate when it comes to detecting Borrelia burgdorferi infection, but many physicians will tell their patients they do not have Lyme disease based solely upon results from these tests. Of these, the PCR is considered the most reliable

Western Blot
- Dr. Crist explains that the Western blot test is usually used as a 'confirmation test' when a patient has already tested positive on one of the above screening tests. He has presented research however (at the 1994 International Lyme Borreliosis Conference held in Bologna, Italy) that shows the Western blot to be much more accurate, detecting Bb infection when a patient has tested negative on previous screening tests. The Western blot test is a sensitive method of detecting antibodies to Bb in a patients blood by forcing them through a gel containing Bb fragments.

Recommended Labs: IGeneX , Medical Diagnostic Laboratories

Bowen Q-RiBb (Quantitative Rapid Identification of Borrelia Burgdorferi)
- A new testing method, that potentially represents a major breakthrough, is the Q-RiBb test, which has now been approved a US patent.

Developed by Jo Anne Whitaker, M.D., a prominent international medical researcher, and Lyme disease patient, the Q-RiBb test is unique in its approach and potentially offers much greater accuracy than conventional testing. The method uses a fluorescent antibody technique on whole blood. As it is "quantative", the test can determine the extent of infection and may therefore be able to distinguish the carriers from the patients with serious disease.

Another advantage is the brief time required to complete the test.
A preliminary report of the findings is provided within 24 hours of receiving the specimen and the final report includes digital photographs of the findings. Finally, the Q-RiBb is the only test that is unaffected by whether the patient is currently (or recently has been) taking antibiotics.

Bowen Research & Training Institute

Flow Cytometry
- This is the test that Central Florida Research Inc is now using to test for Borrelia burgdorferi infection and Lyme disease. It is considered by many to be the "gold standard" of diagnostic testing at present. The test is essentially a refinement of the Q-RiBb and is such is much more accurate than the Western Blot, and of course the other screening methods.

The Central Florida Research Inc website states that a "A Borrelia burgdorferi fluorescent antibody is used to detect the antigen in whole blood. The test is set up manually and read by Flow Cytometry. The Flow Cytometer can count the number of organisms in 100,000 events in 2 minutes and 50,000 in 1 minute. To visually count the organisms in 100,000 events or 50,000 events using a microscope would be almost an impossibility.

The Flow Cytometer
counts the number of all events passing through the aperture and enumerates the organisms that react with the antibody. The test result will be reported as a percent of the counted events." Essentially this statement is explaining that the Flow Cytometry test is quantitative and is able to detect the severity of infection rather than simply giving a positive or negative result for the presence of Bb. For more information visit the Central Florida Research Inc website.

Central Florida Research Inc

Erythema Migrans Biopsy

If erythema migrans rash is present then a biopsy should be carried out as soon as possible. If Borrelia burgdorferi is present then it can be cultured from a tissue biopsy. This procedure can give a crucial early diagnosis of Lyme disease many weeks before antibodies will show up on many of the commonly used laboratory tests.

Testing Conclusions

Most Lyme experts agree that current testing methods (with the possible exception of the Q-RiBb) are less than precise and should not be the sole tool used for diagnosing Lyme disease. Instead, a thorough review of possible exposure to ticks, patient symptoms, biopsy, and then the more accurate diagnostic tests, PCR and Western blot, should be used in combination to make a Lyme disease diagnosis.

Treatment

If Lyme disease is diagnosed in its early stages (erythema migrans), it can often be treated quickly and successfully with oral antibiotic treatment. If there are other symptoms, the patient may be admitted to hospital for further investigation and possible further treatment with antibiotics. In cases of chronic Lyme disease a more holistic approach is often required to restore the patients health. In these cases functional/integrative and alternative medicine approaches are arguably more successful.

Standard Antibiotic Drug Therapy

Medicines used against Lyme disease include:

Oral: doxycycline (eg Vibramycin) (except in children), amoxicillin (eg Amoxil) or cephalosporin antibiotics are the usual first choices.

Studies have demonstrated that when Lyme disease is caught early, 4-6 weeks of oral antibiotics results in remission and apparent cure in most but not all cases. Several months is not uncommon and may be necessary in many instances before a cure is achieved. It is particularly important in this instance to regularly test blood levels on the antibiotic to make sure the level required to eradicate the Bb infection is maintained.

Injection:
benzylpenicillin (eg Crystapen), cefotaxime (Claforan) and ceftriaxone (Rocephin) are the usual choices.

Used mainly when severe localized infection is present, especially when there is brain or heart involvement.

Alternative Treatments

Although physicians practicing integrative/functional medicine try to avoid the use of antibiotic drugs because of their potential side-effects, when it comes to treating Lyme disease they often resort to these medicines. The difference is that they will also use other therapies concurrently in an effort bolster resistance to the infection naturally.

Steven J. Bock, MD speaks about the integrative treatment of Lyme disease in the International Journal of Integrative Medicine (May/June 1999):

"Patients with Lyme disease are placed on a nutritional regimen that includes anti-inflammatory eicosanoids, such as fish oil and borage seed oil. A high potency multivitamin/mineral formula is also used. Since muscle pain and spasm are present in many cases, a calcium/magnesium supplement is usually prescribed.

Extra magnesium is recommended if symptoms are predominantly of a fibromyalgia symptoms are secondary to the underlying disease. CoQ10 and other mitochondrial nutrients (e.g., carnitine and lipoic acid) promote energy production. Intravenous nutrients, such as vitamin C and B vitamins, are often utilized for immune function enhancement.

When a patient is placed on antibiotic therapy, it is imperative to give him or her probiotics (e.g., Lactobacillus, acidophilus or bifidum) and Saccharomyces boulardii. this prevents imbalance in the intestinal flora, which could lead to intestinal dysbiosis and/or C.dificile infection.

Chronic candidiasis and intestinal dysbiosis are frequently encountered in the treatment of Lyme patients. In some cases, natural anti-fungal therapy is utilized. Nystatin or fluconazole can also be used. Occasionally, intestinal cleansing is necessary. milk thistle extract can help prevent potential dysfunction of liver enzymes from antibiotic therapy."

Other alternative treatments that may benefit the Lyme disease patient include acupuncture, stress reduction techniques and nutritional support for cognitive dysfunction, such as Gingko Biloba, L-Acetyl-Carnitine and pregnenolone.

Dr. Whitaker, who developed the Q-RiBb test, has also developed a treatment called the Bowen Technique. According to www.bowen.org " The Bowen- ANS (Autonomic Nervous System ) technique uses a series of simple gentle moves across muscle and connective tissue. This process is very effective in helping balance many muscular, skeletal nerve or chronic conditions. By balancing the parasympathetic and sympathetic portions of the Autonomic Nervous System, the body begins to heal itself and to be relieved of toxins.

Bowen Therapy is covered by Medicare and many insurances."

Samento
- An exciting new natural antibacterial for the treatment of Lyme disease. Samento is also known as TOA-free Cat's Claw or Pentacyclic Alkaloid Chemotype Uncaria tomentosa, to give it it's scientific name. This rare form of the Cat's Claw plant contains pentacyclic oxindole alkaloids (POAs) that have powerful immune system modulating properties, especially relating to non-specific and cellular immunity. What makes Samento special is that it does not contain tetracyclic oxindole alkaloids (TOAs) that, even in very small amounts, inhibit this immune modulating action. Samento also contains generous amounts of acid glycosides, the antimicrobial chemicals found in the latest quinolone antibiotic drugs, the standard treatment for Lyme disease.

The combination of POAs and acid glycosides appears to be highly effective in treating Lyme disease by directly killing off the Borrelia burgdorferi spirochete, and increasing the patients immune response and hence resistance, to the pathogen.

In an 8 month pilot study, 28 patients suffering from chronic Lyme disease, who had all tested positive for Borrelia burgdorferi on the Western blot test, were split into 2 groups. The control group received standard antibiotic therapy and the experimental group was given Samento. At the end of the study, of the 14 patients in the control group, 3 improved slightly, 3 got worse and the rest had no change. In the Samento group, 85% tested negative for Borrelia burgdorferi when the Western blot was repeated, and all the patients reported a large improvement in their condition.

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