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NON-TICK TRANSMISSION Note: Visit the FOOD SUPPLY section on Dairy Cows, Cattle and Livestock for additional examples of Non-Tick Transmission to their Offspring, and their role as Carriers/Reseroirs, including how this affects our Food Supply.
Transmission through Urine White Footed Mouse The prevalence and significance of Borrelia burgdorferi in the urine of feral reservoir hosts
Live Borrelia burgdorferi were isolated from the blood and/or urine of white-footed mice (Peromyscus leucopus) collected on Shelter Island, New York, in 1984 and 1985.
Prevalence of spirochetes in urine was consistently higher than in blood or both fluids simultaneously. Spirochetes remained viable for 18-24 hours in urine and were maintained in culture for one week.
Mice removed from the field were spirocheturic for at least 13 months. One spirocheturic mouse developed spirochetemia one month after field removal indicating the pathogen can return to the peripheral circulation.
Twenty-one kidneys from 22 mice had spirochetes in the interstitial areas and bridging the tubules. A positive correlation between Babesia microti infection and spirocheturia was seen. Although the mechanism of entry into the urine is unknown, B. microti infection may increase glomerular permeability.
Babesia induced hematuria may provide possible nutrients to maintain spirochetes. Urine may provide a method for contact non-tick transmission of B. burgdorferi in natural rodent populations particularly during periods of nesting and/or breeding.
Beagles Clinical and serologic evaluations of induced Borrelia burgdorferi infection in dogs
Adult Beagles were used to evaluate clinical signs and serologic response after inoculation with, or exposure to, Borrelia burgdorferi.
An indirect immunofluorescent assay (IFA) and 2 ELISA were used to monitor the serologic response to B burgdorferi. Feeding infected ticks on 4 dogs (group 1) failed to cause seroconversion, and SC inoculation with 500 organisms caused minimal seroconversion in 2 of 4 dogs (group 2).
At 56 days, approximately 3.01 X 10(8) B burgdorferi organisms were injected IV into group-1 dogs, and intraperitoneally into group-2 dogs. A control group of 4 dogs (group 3) had noninfected ticks feed on them, and then were given IV injection of physiologic saline solution. Increases in immunoglobulin M (IgM) titers were detected in 2 of 4 group-2 dogs approximately 7 days after the initial exposure. These titers returned to negligible values 20 days later.
Immunoglobulin G titers increased approximately 10 days after the initial exposure and were mildly increased 56 days later, when dogs were exposed a second time. Both the IV and intraperitoneal injections (second exposures) resulted in increased IgM titers, which in both groups eventually returned to preexposure values after approximately 2 months.
Immunoglobulin G titers increased within a week after the second exposure, and in 3 dogs monitored for 8 months, returned to negligible values after the 8-month period. One control dog had a slightly increased IgG titer 24 days after the second inoculation. The possibility of urine transmission is suggested.
Clinical status, hemograms, serum biochemical profiles, ECG and results of urinalyses remained normal throughout the study.
Transmission through Contact Beagles
Contact Transmission
Subclinical
Persistent Infections Experimental inoculation of dogs with Borrelia burgdorferi
To determine if dogs could serve as a reservoir for Borrelia burgdorferi, three beagles were inoculated subcutaneously (SQ) with 200 laboratory cultured spirochetes which were originally isolated from blood of a Peromyscus leucopus from Ft. McCoy, Wisc. One four month old beagle was inoculated SQ with 5 ground Ixodes dammini from Shelter Island, N.Y.
which came from an area with a 50% B. burgdorferi tick infection rate; and another uninfected four month old beagle was housed loose on the floor with the tick inoculated dog. All three spirochete inoculated beagles developed IFA antibody titers to B. burgdorferi of (7 log2) to (8 log2) by day 28 post inoculation. All were apparently healthy and no spirochetes were cultured from the blood.
In an attempt to exacerbate the disease two of the dogs were given 3 mg of dexamethasone on day 68 post inoculation. B. burgdorferi was isolated from blood of all these dogs on days 4 and 97 days post inoculation.
The tick inoculated dog developed a B. burgdorferi IFA antibody titer of (10 log2) by day 14 post inoculation. The contact exposed dog also developed a B. burgdorferi IFA antibody titer of (7 log2) on post contact day 21 indicating contact infection. B. burgdorferi was not isolated from either of these dogs.
These results indicate that, contact transmission of B. burgdorferi may occur between dogs, dogs can be subclinically infected with B. burgdorferi and have persistent infections.
Transmission to Offspring Mice Transmission to offsprings
B. burgdorferi can be transmited by other modes besides the tick bite Transmission of Borrelia burgdorferi from Experimentally Infected Mating Pairs to Offsprings in a Murine Model
Abstract:
The current literature on B. burgdorferi=s mode of transmission in animal models supports only transmission of the organism by an infected tick bite. In an effort to develop a murine model for studying other modes of transmission of B. burgdorferi, we started with the well studied C3H/HeJ mouse. Abzug et al. has shown that splenectomy increases the in utero transmission of enteroviruses in a murine model.
Splenectomized 6-8 week-old mice were divided into 4 groups. Groups A, B, and C had 23, 24, and 26 mating pairs respectively. Immediately prior to mating, in group A only females, in group B only males, and in group C both females and males were infected subcutaneously with 106-107 B. burgdorferi in 250 ml SKB II media. The control group D had 12 mating pairs in which both male and females received sterile SKB II as placebo. The resulting pups were sacrificed at 1, 7, 14, and 21 days of age.
The milk content of the stomach, sections from ear, skin, heart, liver, spleen, brain, bladder, and kidney of the 1, 7, and 14 day-old pups were cultured for B. burgdorferi. The cultures were read at 3, 6, and 9 weeks post incubation. The above mentioned tissues except milk were also cultured from sacrificed 21 day-old weanlings.
Transmission to offsprings was indicated when B. burgdorferi was isolated from any tissue from a given pup. From the experimentally infected females in which the milk was cultured ( total 25 females in groups A and C), 2 (8%) transmitted B. burgdorferi to their pups on day one via their milk: 2 pups from a litter of 4 in group C, 1 pup from a litter of 8 in group A. No transmission was detected via milk on days 7 or 14. Among 49 infected females from groups A and C, 5 (10.2%) transmitted B. burgdorferi to their pups either in utero or intrapartum.
Two of the transmissions were detected on day 1 ( litter one, 2/6; litter two, 1/7), two on day 7 (litter three, 1/7; litter four, 2/6), and one on day 14 (litter five, 2).
Interestingly, four of the litters from the mating pairs in group B had infected pups (litter one, 3/5; litter two, 1/8; litter three, 3/6; and litter four, 2/4). These results indicate that B. burgdorferi can be transmited by other modes besides the tick bite in speenectomized mice.
The described mouse model with further modifications may provide a tool for studying such transmission modes and treatment strategies.
Transmission by Semen Semen
Animal Semen Survive in cryopreservation Viability of Borrelia burgdorferi in stored semen
Semen from 5 dogs, 3 bulls and 3 rams were collected ... divided in 2 equal parts, BSK-H medium and B. burgdorferi added + 4 controls.
A. was stored at 5 degr. C for 48 hours
B. was cryopreserved in liquid nitrogen at -196 degr. C for 12 weeks The specimens were thawed in waterbath to 37 degr. C.
A drop on object glass was examined in phase contrast microscope for sperm motility and viable / motile B. burgdorferi, graded on a scale 0-3.
A significant amount of spirochetes survived cryopreservation and no significant different found between the 2 different storage methods or between the 3 animal species.
The viability of B. burgdorferi in cryopreservation was better than spermatozoa.
The second case described is that of a woman who developed Lyme borreliosis symptoms, when intestinal content of an infected tick came into contact with her conjunctiva. In both cases the diagnosis is based on clinical picture and positive serological tests.
The first case shows the probability of contracting Lyme borreliosis when the duration of the tick's attachment to the skin is less than 24 hours. The second case, described demonstrates transmission of B. burgdorferi by contact. Apart from the two cases mentioned in the abstract the author tells about two other cases of direct transmission, he'd heard of:
1. 1989 Prag conference - a laboratory worker, during handling of a tick, the tick dropped onto a hot lamp. The tick bursted and the intestinal material hit the eye of the lab worker, who developed conjunctivitis and positive borrelia serology.
If anyone know the identity and country of this lab. worker - then please send a note to me at [email protected] 2. A case from Sweden: EM developed in a scratch-wound that was contaminated with maneuver in which B. burgdorferi was found.
To determine if dogs could serve as a reservoir for Borrelia burgdorferi, three beagles were inoculated subcutaneously (SQ) with 200 laboratory cultured spirochetes which were originally isolated from blood of a Peromyscus leucopus from Ft. McCoy, Wisc.
One four month old beagle was inoculated SQ with 5 ground Ixodes dammini from Shelter Island, N.Y. which came from an area with a 50% B. burgdorferi tick infection rate; and another uninfected four month old beagle was housed loose on the floor with the tick inoculated dog.
All three spirochete inoculated beagles developed IFA antibody titers to B. burgdorferi of (7 log2) to (8 log2) by day 28 post inoculation. All were apparently healthy and no spirochetes were cultured from the blood.
In an attempt to exacerbate the disease two of the dogs were given 3 mg of dexamethasone on day 68 post inoculation. B. burgdorferi was isolated from blood of all these dogs on days 4 and 97 days post inoculation.
The tick inoculated dog developed a B. burgdorferi IFA antibody titer of (10 log2) by day 14 post inoculation. The contact exposed dog also developed a B. burgdorferi IFA antibody titer of (7 log2) on post contact day 21 indicating contact infection. B. burgdorferi was not isolated from either of these dogs.
These results indicate that, contact transmission of B. burgdorferi may occur between dogs, dogs can be subclinically infected with B. burgdorferi and have persistent infections.
Clinical and serologic evaluations of induced Borrelia burgdorferi infection in dogs. Greene RT, Levine JF, Breitschwerdt EB, Walker RL, Berkhoff HA, Cullen J, Nicholson WL. Am J Vet Res 1988 Jun; 49(6): 752-7
Adult Beagles were used to evaluate clinical signs and serologic response after inoculation with, or exposure to, Borrelia burgdorferi. An indirect immunofluorescent assay (IFA) and 2 ELISA were used to monitor the serologic response to B burgdorferi.
Feeding infected ticks on 4 dogs (group 1) failed to cause seroconversion, and SC inoculation with 500 organisms caused minimal seroconversion in 2 of 4 dogs (group 2). At 56 days, approximately 3.01 X 10(8) B burgdorferi organisms were injected IV into group-1 dogs, and intraperitoneally into group-2 dogs.
A control group of 4 dogs (group 3) had noninfected ticks feed on them, and then were given IV injection of physiologic saline solution. Increases in immunoglobulin M (IgM) titers were detected in 2 of 4 group-2 dogs approximately 7 days after the initial exposure. These titers returned to negligible values 20 days later.
Immunoglobulin G titers increased approximately 10 days after the initial exposure and were mildly increased 56 days later, when dogs were exposed a second time. Both the IV and intraperitoneal injections (second exposures) resulted in increased IgM titers, which in both groups eventually returned to preexposure values after approximately 2 months.
Immunoglobulin G titers increased within a week after the second exposure, and in 3 dogs monitored for 8 months, returned to negligible values after the 8-month period. One control dog had a slightly increased IgG titer 24 days after the second inoculation. The possibility of urine transmission is suggested.
Clinical status, hemograms, serum biochemical profiles, ECG and results of urinalyses remained normal throughout the study.(ABSTRACT TRUNCATED AT 250 WORDS)
The prevalence and significance of Borrelia burgdorferi in the urine of feral reservoir hosts. Bosler EM, Schulze TL. Zentralbl Bakteriol Mikrobiol Hyg A 1986 Dec; 263(1-2): 40-4
Live Borrelia burgdorferi were isolated from the blood and/or urine of white-footed mice (Peromyscus leucopus) collected on Shelter Island, New York, in 1984 and 1985. Prevalence of spirochetes in urine was consistently higher than in blood or both fluids simultaneously.
Spirochetes remained viable for 18-24 hours in urine and were maintained in culture for one week. Mice removed from the field were spirocheturic for at least 13 months.
One spirocheturic mouse developed spirochetemia one month after field removal indicating the pathogen can return to the peripheral circulation. Twenty-one kidneys from 22 mice had spirochetes in the interstitial areas and bridging the tubules.
A positive correlation between Babesia microti infection and spirocheturia was seen. Although the mechanism of entry into the urine is unknown, B. microti infection may increase glomerular permeability.
Babesia induced hematuria may provide possible nutrients to maintain spirochetes. Urine may provide a method for contact non-tick transmission of B. burgdorferi in natural rodent populations particularly during periods of nesting and/or breeding.
Transmission by MILK or food?
Most spirochetes (and other bacteria) ingested will probably be killed by the high acidic content in the stomach, but people with achlorhydria and newborns that have very low stomach acid production, does not have this protective barrier and might be at increased risk for getting infected by the oral route, if they ingest live spirochetes.
Pasteurizing the milk and never eat semi-raw meat - must be recommended as prophylaxis.
Borrelia burgdorferi infection in Wisconsin horses and cows. Burgess EC. Ann N Y Acad Sci 1988; 539: 235-43
Blood samples from Wisconsin horses and cows suspected of having clinical disease due to Borrelia burgdorferi infection were submitted by veterinary practitioners.
All serum, milk, colostrum, and synovial samples were tested for B. burgdorferi antibodies by immunofluorescence. Whole blood, milk, colostrum, and synovial fluid samples were cultured for B. burgdorferi.
Records were kept on the clinical signs of antibody-positive animals, date of sample, and location of the animal by county. Of the samples tested for antibodies 282/430 cow sera, 118/190 horse sera, 5/10 cow synovial fluids, 3/6 horse synovial fluids, 2/3 cow colostrums, 0/44 cow milk samples and 1 aborted fetus serum were antibody positive at a titer of 1:128 or greater.
Of samples cultured 7/156 cow bloods, 2/35 horse bloods, 1/14 cow synovial fluids, 0/4 synovial fluids, 1/3 cow colostrums, 0/44 cow milk, and 2/10 cow urine samples were B. burgdorferi culture positive. For both cows and horses October and May were the two peak months for the number of antibody-positive samples.
The most frequent clinical signs in antibody-positive horses and cows were lameness and swollen joints, but many also had stiffness, laminitis, abortions, and fevers. Not all antibody-positive animals showed clinical signs.
These findings show that B. burgdorferi infection occurs in horses and cows and can cause clinical illness in some but not all animals. Infection in cows and horses occurs most frequently 1 month after the emergence of adult I. dammini.
Because spirochetes could be isolated from blood, synovial fluid, colostrum, and urine, these animals could be important in providing an infected blood meal for ticks and bringing B. burgdorferi in direct contact with humans.
Borrelia burgdorferi: another cause of foodborne illness?
Farrell GM, Marth EH. Int J Food Microbiol 1991 Dec; 14(3-4): 247-60
Borrelia burgdorferi was identified as the etiological agent of Lyme disease in 1982. This Gram-negative spirochete is classified in the order Spirochaetales and the family Spirochaetaceae.
The pathogen is fastidious, microaerophilic, mesophilic and metabolises glucose through the Embden-Meyerhof pathway. A generation time of 11 to 12 h at 37 degrees C in Barbour-Stoenner-Kelly medium has been reported.
Lyme disease, named after Lyme in Connecticut, is distributed globally. It is the most commonly reported vector-borne disease in the United States, where the incidence is highest in the eastern and midwestern states.
Since establishment of national surveillance in 1982, there has been a nine-fold increase in the number of cases reported to the U.S. Centers for Disease Control. The deer tick of the genus Ixodes is the primary vector of Lyme borreliosis. The tick may become infected with B. burgdorferi, by feeding on an infected host, at any point in its 2-year life cycle which involves larval, nymphal and adult stages.
The infection rate in deer ticks may be as high as 40% in endemic areas. The primary vertebrate reservoirs for Ixodes are the white-footed mouse (Peromyscus leucopus) and the white-tailed deer (Odocopileus virginianus).
Dairy cattle and other food animals can be infected with B. burgdorferi and hence some raw foods of animal origin might be contaminated with the pathogen. Recent findings indicate that the pathogen may be transmitted orally to laboratory animals, without an arthropod vector.
Thus, the possibility exists that Lyme disease can be a food infection. In humans, the symptoms of Lyme disease, which manifest themselves days to years after the onset of infection, may involve the skin, cardiac, nervous and/or muscular systems, and so misdiagnosis can occur.
Evidence for in utero Transmission of Borrelia burgdorferi from Naturally Infected Cows Journal of Spirochetal and Tick-Borne Diseases 1998; 5(4):54-62
Leibstein MM, Khan MI, Bushmich SL (excerpts from the abstract:
Five of 15 adult cows were spirochetemic at parturition; 4 of the calves from these cows were also spirochetemic at birth (PCR).
Spirochetes were cultured from the placentas in 2 of 10 cows and from the uterine fluid in 1 of 8 cows. Borrelia burgdorferi DNA was detected in the colostrum in 4 of 12 cows. Three of 15 calves were stillborn; Borrelia burgdorferi DNA was detected by PCR in 3 of 3 and spirochetes cultured from 2 of 3 stillborn calves.
Fetal tissues from which Borrelia burgdorferi DNA was detected include blood, spleen, bladder, kidney, synovial fluid and tissue, heart, cerebrum, and aqueous humor. Borrelia burgdorferi was cultured from the spleen of one stillborn calf and the kidney of another.
Detection of Borrelia burgdorferi DNA from the tissues of stillborn calves, as well as spirochetemia in neonatal liveborn and stillborn calves, gives evidence for in utero transmission of Borrelia burgdorferi in naturally infected dairy cattle.)
Last, a splenectomized mice study, yet unpublished, but Sousan Altaie very kindly provided me with her draft text, and she has allowed me to refer to her data, as stated to me in a mail per 28-03-00:
Marie, ... If you like you may refer to my work as personal communication, unpublished data. You may also refer to the two published abstracts. ... Susan Transmission of Borrelia burgdorferi from Experimentally Infected Mating Pairs to Offsprings in a Murine Model.
Altaie SS, Mookherjee S, Assain A, AL-TAIE F, Nakeeb SM, SAEEDA Y. Siddiqui SY Departments of Pediatrics and Pathology, State University of New York at Buffalo and The Children's Hospital of Buffalo, NY. abstract:
The current literature on B. burgdorferi=s mode of transmission in animal models supports only transmission of the organism by an infected tick bite. In an effort to develop a murine model for studying other modes of transmission of B. burgdorferi, we started with the well studied C3H/HeJ mouse.
Abzug et al. has shown that splenectomy increases the in utero transmission of enteroviruses in a murine model.
Splenectomized 6-8 week-old mice were divided into 4 groups. Groups A, B, and C had 23, 24, and 26 mating pairs respectively. Immediately prior to mating, in group A only females, in group B only males, and in group C both females and males were infected subcutaneously with 106-107 B. burgdorferi in 250 ml SKB II media.
The control group D had 12 mating pairs in which both male and females received sterile SKB II as placebo. The resulting pups were sacrificed at 1, 7, 14, and 21 days of age. The milk content of the stomach, sections from ear, skin, heart, liver, spleen, brain, bladder, and kidney of the 1, 7, and 14 day-old pups were cultured for B. burgdorferi.
The cultures were read at 3, 6, and 9 weeks post incubation. The above mentioned tissues except milk were also cultured from sacrificed 21 day-old weanlings. Transmission to offsprings was indicated when B. burgdorferi was isolated from any tissue from a given pup.
From the experimentally infected females in which the milk was cultured ( total 25 females in groups A and C), 2 (8%) transmitted B. burgdorferi to their pups on day one via their milk: 2 pups from a litter of 4 in group C, 1 pup from a litter of 8 in group A. No transmission was detected via milk on days 7 or 14. Among 49 infected females from groups A and C, 5 (10.2%) transmitted B. burgdorferi to their pups either in utero or intrapartum.
Two of the transmissions were detected on day 1 ( litter one, 2/6; litter two, 1/7), two on day 7 (litter three, 1/7; litter four, 2/6), and one on day 14 (litter five, 2). Interestingly, four of the litters from the mating pairs in group B had infected pups (litter one, 3/5; litter two, 1/8; litter three, 3/6; and litter four, 2/4). These results indicate that B. burgdorferi can be transmited by other modes besides the tick bite in speenectomized mice.
The described mouse model with further modifications may provide a tool for studying such transmission modes and treatment strategies.
This study has also been presented on the following conferences, but I don't have the abstracts (I any of you have them, please send me a copy, will you?):
Sayahtaheri Altaie S, Assian E, Mookherjee S, Al-Taie F, et al. 1997. Transplacental Transmission of Borrelia burgdorferi in a Murine Model. Abstract, p. 34.
10th Annual International Scientific Conference on Lyme Disease & other Tick-borne Disorders. Lyme Disease Foundation.
Sayahtaheri Altaie S, Assian E, Mookherjee S, Al-Taie F, et al. 1997. Transplacental Transmission of Borrelia burgdorferi in a Murine Model. Abstract D-130, p. 231. 97th Annual Meeting of the American Society for Microbiology.
of borreliosis in humans have never been proved. AFAIK it haven't been investigated (?), nor has it been investigated if male's excrete spirochaetae in semen (?), if this could be the case, semen could also be a route of transmission - wouldn't in fact be surprising as it's 'cousin' syphilis is sexually contagious.
In above study by S. Altaie et al, sexual transmission could not be demonstrated between mating pairs of splenectomized mice.
Viability of Borrelia burgdorferi in stored semen. Kumi Diaka J, Harris O. Br Vet J 1995 Mar-Apr; 151(2): 221-4
no abstract Semen from 5 dogs, 3 bulls and 3 rams were collected ... divided in 2 equal parts, BSK-H medium and B. burgdorferi added + 4 controls.
A was stored at 5 degr. C for 48 hours B was cryopreserved in liquid nitrogen at -196 degr. C for 12 weeks The specimens were thawed in waterbath to 37 degr. C.
A drop on object glass was examined in phase contrast microscope for sperm motility and viable / motile B. burgdorferi, graded on a scale 0-3.
A significant amount of spirochetes survived cryopreservation and no significant different found between the 2 different storage methods or between the 3 animal species.
The viability of B. burgdorferi in cryopreservation was better than spermatozoa
Exchange of Borrelia burgdorferi between Ixodes persulcatus (Ixodidae:Acarina) sexual partners.Alekseev AN, Dubinina HV. Zoological Institute of Russian Academy of Sciences, Universitetskaja nab., St. Petersburg, Russia.
Borrelia burgdorferi sensu lato infection rate in Ixodes persulcatus Schulze maintained at different relative humidity gradients in male and females pairs, separated by sex, and in ticks of both sexes having either normal or abnormal exoskeleton were compared.
Ticks were collected in the St. Petersburg Region of Russia during 1992 and 1994. We observed that the infection rate among the ticks maintained as sexual pairs was 1.75-2.00 times higher than that among ticks maintained singles, indicating a borreliae interchange between sexual partners. This pathogen interchange was thought to result from a venereal or omovampiric (cannibalistic) mode of borreliae transmission.
Borrelia burgdorferi s.l. was determined to be present in 22.9% (112 infected specimens of 489 total), whereas infection occurred in 17.4% of single females and 16.5% of single males. The data indicate the importance of isolating ticks sexually during quantitative disease investigations with borreliae as well as tick-borne encephalitis virus and other tick-borne pathogens.
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