Bartonella bovis and Bartonella chomelii infection in dairy cattle and their ectoparasites in Algeria.

Bartonella are blood-borne and vector-transmitted bacteria, some of which are zoonotic. B. bovis and B. chomelii have been reported in cattle. However, no information has yet been provided on Bartonella infection in cattle in Algeria. Therefore, 313 cattle from 45 dairy farms were surveyed in Kabylia, Algeria, in order to identify Bartonella species infecting cattle using serological and molecular tests. In addition, 277 ticks and 33 Hippoboscidae flies were collected. Bartonella bovis and B. chomelii were identified as the two species infecting cattle. Bartonella DNA was also amplified from 6.8 % (n = 19) of ticks and 78.8 % (n = 26) of flies. Prevalence of B. bovis DNA in dairy cattle was associated both with age and altitude. This study is the first one to report of bovine bartonellosis in Algeria, both in dairy cattle and in potential Bartonella vectors, with the detection of B. bovis DNA in tick samples and B. chomelii in fly samples.

Conjunctival bacterial and fungal flora and cutaneous fungal flora in healthy domestic rabbits (Oryctolagus cuniculus).

OBJECTIVES: To determine the normal conjunctival bacterial and fungal flora in healthy rabbits, to compare the fungal flora from the conjunctival sac with periocular cutaneous fungal flora and to evaluate potential changes in type of micro-organisms over time. MATERIALS AND METHODS: Samples from 50 pet rabbits with a normal ophthalmic examination were obtained from conjunctival sacs using sterile cotton swabs and from the interpupillary skin space using a piece of sterile carpet. Samples were collected twice, 4 weeks apart, and cultured to evaluate bacterial and fungal populations. RESULTS: Bacteria were recovered from 35.9% of conjunctival swabs. Corynebacterium species (32.4%), Staphylococcus species (18.3%), Pasteurella species (9.9%) and Moraxella species (8.5%) were the most frequently isolated organisms. Fungi were recovered from 14.6% of conjunctival swabs. The most common fungal genera were Aspergillus (37%), Scopulariopsis (14.8%) and Penicillium (11.1%). Fungal cultures from periocular integument yielded positive results in 96.3% of samples; the same fungal organism was isolated in both conjunctival and cutaneous culture in nine of 20 rabbits (45%). When comparing results 4 weeks apart in 32 of 50 animals, only three rabbits with a positive bacterial culture and one rabbit with a positive fungal culture had the same result at the subsequent test. CLINICAL SIGNIFICANCE: Fungi and bacteria isolated from the conjunctival sac of rabbits may represent transient seeding from the environment. To our knowledge, this is the first report of fungal composition of the conjunctival sac of rabbits and evaluation of the potential transient nature of both bacterial and fungal composition of this conjunctival flora.

Clinical description of Corynebacterium urealyticum urinary tract infections in 11 dogs and 10 cats.

OBJECTIVES: To examine the clinical signs, laboratory findings, possible predisposing factors, antimicrobial sensitivity and outcomes of dogs and cats diagnosed with urinary tract infection caused by Corynebacterium urealyticum. MATERIALS AND METHODS: Retrospective, observational study of dogs and cats that were diagnosed with urinary tract infection caused by C. urealyticum. RESULTS: This report concerns a total of 11 dogs and 10 cats. All cats had a history of urethral catheterisation, and six cats had undergone urological surgery before diagnosis of this specific infection. Dogs had history of neurogenic urinary incontinence (n=5), urethral catheterisation (n=4), urological surgery (n=4), lower urinary tract disease (n=2) or prostatic disease (n=1). In seven animals, previous urine culture was negative. Median urine pH was 8.6 (interquartile range 7.5 to 9), and 19 animals had pyuria and struvite crystalluria. Encrusted cystitis was diagnosed by abdominal ultrasound in nine animals. All but one isolate were resistant to at least three antimicrobial classes. Thirteen isolates were sensitive to tetracyclines. There was bacterial and clinical resolution of the infection in nine dogs and seven cats. Two animals died from sepsis and two cats were euthanased because of clinical deterioration. CLINICAL SIGNIFICANCE: Urinary tract infection caused by C. urealyticum should be suspected in animals with pre-existing urinary disorders, especially if urine is alkaline and there is struvite crystalluria, negative routine urine culture and encrusted cystitis.

Bovine anaplasmosis and tick-borne pathogens in cattle of the Galapagos Islands.

A cross-sectional study was conducted to determine the species of Anaplasma spp. and estimate its prevalence in cattle of the three main cattle-producing Galapagos Islands (Santa Cruz, San Cristóbal and Isabela) using indirect PCR assays, genetic sequencing and ELISA. Ticks were also collected from cattle and scanned for 47 tick-borne pathogens in a 48 × 48 real-time PCR chip. A mixed effects logistic regression was performed to identify potential risk factors explaining Anaplasma infection in cattle. A. phagocytophilum was not detected in any of the tested animals. Genetic sequencing allowed detection of A. platys-like strains in 11 (36.7%) of the 30 Anaplasma spp.-positive samples analysed. A. marginale was widespread in the three islands with a global between-herd prevalence of 100% [89; 100]95% CI and a median within-herd prevalence of 93%. A significant association was found between A. marginale infection and age with higher odds of being positive for adults (OR = 3.3 [1.2; 9.9]95% Bootstrap CI ). All collected ticks were identified as Rhipicephalus microplus. A. marginale, Babesia bigemina, Borrelia theileri and Francisella-like endosymbiont were detected in tick pools. These results show that the Galapagos Islands are endemic for A. marginale.

High Prevalence of Anaplasma spp. in Small Ruminants in Morocco.

The prevalence of infection by Anaplasma spp. (including Anaplasma phagocytophilum) was determined using blood smear microscopy and PCR through screening of small ruminant blood samples collected from seven regions of Morocco. Co-infections of Anaplasma spp., Babesia spp, Theileria spp. and Mycoplasma spp. were investigated and risk factors for Anaplasma spp. infection assessed. A total of 422 small ruminant blood samples were randomly collected from 70 flocks. Individual animal (breed, age, tick burden and previous treatment) and flock data (GPS coordinate of farm, size of flock and livestock production system) were collected. Upon examination of blood smears, 375 blood samples (88.9%) were found to contain Anaplasma-like erythrocytic inclusion bodies. Upon screening with a large spectrum PCR targeting the Anaplasma 16S rRNA region, 303 (71%) samples were found to be positive. All 303 samples screened with the A. phagocytophilum-specific PCR, which targets the msp2 region, were found to be negative. Differences in prevalence were found to be statistically significant with regard to region, altitude, flock size, livestock production system, grazing system, presence of clinical cases and application of tick and tick-borne diseases prophylactic measures. Kappa analysis revealed a poor concordance between microscopy and PCR (k = 0.14). Agreement with PCR is improved by considering microscopy and packed cell volume (PCV) in parallel. The prevalence of double infections was found to be 1.7, 2.5 and 24% for Anaplasma-Babesia, Anaplasma-Mycoplasma and Anaplasma-Theileria, respectively. Co-infection with three or more haemoparasites was found in 1.6% of animals examined. In conclusion, we demonstrate the high burden of anaplasmosis in small ruminants in Morocco and the high prevalence of co-infections of tick-borne diseases. There is an urgent need to improve the control of this neglected group of diseases.

Short Report: Identification of a Potential Marker of Anaplasma Phagocytophilum Associated with Cattle Abortion.

Anaplasma phagocytophilum is a tick-borne pathogen that causes tick-borne fever in domestic ruminants. Tick-borne fever is characterized by diverse symptoms and occasionally causes abortions in domestic ruminants, resulting in significant economic impact. However, in France, the potential frequency of A. phagocytophilum-related abortions is unknown, and thus, it remains difficult to estimate the full extent of its economic impact. This gap in our knowledge is likely explained, at least in part, by the absence of suitable and specific tools capable of detecting A. phagocytophilum associated with abortion. Our objective was to identify a genetic marker able to differentiate A. phagocytophilum strains isolated from domestic ruminants that had aborted compared to those that had not. Thus, we typed a total of 123 A. phagocytophilum obtained from cattle, of which 25 originated from cows that had aborted, via multiple-locus variable-number tandem repeat (VNTR) analysis. These included 56 new A. phagocytophilum samples and 67 previously published A. phagocytophilum samples. A multivariate logistic model demonstrated that the triple-repeat allele of the APV-A VNTR was statistically more frequent in A. phagocytophilum from cattle that had aborted, compared to A. phagocytophilum from cattle that had not aborted (P = 0.03), while controlling for any regional effects (P < 0.0001). For four other VNTR, there were no statistical associations between specific alleles and abortion. The APV-A triple-repeat VNTR allele could thus act as a marker of A. phagocytophilum involved in abortions. If this hypothesis is confirmed in additional samples from other regions, this marker could then be utilized in the development of A. phagocytophilum abortive strain surveillance measures.

Isolation of Bartonella henselae, Bartonella koehlerae subsp. koehlerae, Bartonella koehlerae subsp. bothieri and a new subspecies of B. koehlerae from free-ranging lions (Panthera leo) from South Africa, cheetahs (Acinonyx jubatus) from Namibia and captive cheetahs from California.

Bartonellae are blood- and vector-borne Gram-negative bacteria, recognized as emerging pathogens. Whole-blood samples were collected from 58 free-ranging lions (Panthera leo) in South Africa and 17 cheetahs (Acinonyx jubatus) from Namibia. Blood samples were also collected from 11 cheetahs (more than once for some of them) at the San Diego Wildlife Safari Park. Bacteria were isolated from the blood of three (5%) lions, one (6%) Namibian cheetah and eight (73%) cheetahs from California. The lion Bartonella isolates were identified as B. henselae (two isolates) and B. koehlerae subsp. koehlerae. The Namibian cheetah strain was close but distinct from isolates from North American wild felids and clustered between B. henselae and B. koehlerae. It should be considered as a new subspecies of B. koehlerae. All the Californian semi-captive cheetah isolates were different from B. henselae or B. koehlerae subsp. koehlerae and from the Namibian cheetah isolate. They were also distinct from the strains isolated from Californian mountain lions (Felis concolor) and clustered with strains of B. koehlerae subsp. bothieri isolated from free-ranging bobcats (Lynx rufus) in California. Therefore, it is likely that these captive cheetahs became infected by an indigenous strain for which bobcats are the natural reservoir.

Lack of transplacental transmission of Bartonella bovis.

Transplacental transmission of Bartonella spp. has been reported for rodents, but not for cats and has never been investigated in cattle. The objective of this study was to assess vertical transmission of Bartonella in cattle. Fifty-six cow-calf pairs were tested before (cows) and after (calves) caesarean section for Bartonella bacteremia and/or serology, and the cotyledons were checked for gross lesions and presence of the bacteria. None of the 29 (52%) bacteremic cows gave birth to bacteremic calves, and all calves were seronegative at birth. Neither placentitis nor vasculitis were observed in all collected cotyledons. Bartonella bovis was not detected in placental cotyledons. Therefore, transplacental transmission of B. bovis and multiplication of the bacteria in the placenta do not seem likely. The lack of transplacental transmission may be associated with the particular structure of the placenta in ruminants or to a poor affinity/agressiveness of B. bovis for this tissue.

Proper use of quinolones for canine colitis ambulatory treatment: literature review and REQUEST guidelines.

The literature analysed by REQUEST (REassesing QUinolone European STandard) using inclusion and exclusion criteria allows to make the conclusion that the proper use of fluoroquinolones (FQ) in canine colitis requires rigorously performed qualification to specific antimicrobial treatment. An infectious agent responsive to FQ therapy plays an integral role in the clinical manifestation of canine colitis, especially histiocytic ulcerative colitis (HUC) in young Boxer dogs. This supports the use of FQ in these cases. The Request guidelines for proper use of FQ in canine colitis is established, according to the available literature data.

Experimental infection of horses with Bartonella henselae and Bartonella bovis.

BACKGROUND: Experimental infection of horses with Bartonella species is not documented. OBJECTIVES: Determine clinical signs, hematologic changes, duration of bacteremia, and pattern of seroconversion in Bartonella henselae or Bartonella bovis-inoculated horses. ANIMALS: Twelve (2 groups of 6) randomly selected healthy adult horses seronegative and culture negative for Bartonella spp. METHODS: Experimental/observational study: Group I: B. henselae or saline control was inoculated intradermally into 4 naïve and 2 sentinel horses, respectively. Group II: same design was followed by means of B. bovis. Daily physical examinations, once weekly CBC, immunofluorescent antibody assay serology, real-time polymerase chain reaction (PCR), and twice weekly blood cultures were performed for 6 weeks and at postinoculation day 80 and 139. Bartonella alpha-Proteobacteria growth medium (BAPGM) enrichment blood culture was performed for horses that seroconverted to B. henselae antigens. RESULTS: Transient clinical signs consistent with bartonellosis occurred in some Bartonella-inoculated horses, but hematological alterations did not occur. Three B. henselae-inoculated horses seroconverted, whereas 1 B. bovis-inoculated horse was weakly seropositive. In Group I, B. henselae was amplified and sequenced from BAPGM blood culture as well as a subculture isolate from 1 horse, blood from a 2nd horse, and BAPGM blood culture from a 3rd horse although a subculture isolate was not obtained. All sentinels remained PCR, culture, and serology negative. CONCLUSIONS: Detection of Bartonella sp. in blood after experimental inoculation supports bacteremia and seroconversion. Culture with BAPGM may be required to detect Bartonella sp. Although mild clinical signs followed acute infection, no long-term effects were noted for 2 years postinoculation.

Murine model for Bartonella birtlesii infection: New aspects.

As a model of persistent infection, various aspects of Bartonella birtlesii infection in laboratory mice, including some immunodeficient mice, are presented, particularly focusing on conditions mimicking natural infection. Bacteraemia was explored using different mice strains routes and inoculum doses (3.4-5x10(7)CFU/mouse). Mice became bacteraemic for 5 (C57Bl6/6) to 10 weeks (Balb/c, Swiss) with peaks ranging from 2x10(3) to 10(5)CFU/mL of blood. The ID route induced the most precocious bacteraemia (day 3) while the higher and longer bacteraemia in immunocompetent mice was obtained with SC when infecting Balb/c with approximately 10(3) CFU/mouse. As opposed to ID, SC and IV routes, bacteraemia was obtained with the oral and ocular routes only for high doses (10(7)) and in 33-66% mice. It was significantly higher and longer in CD4-/- mice compared to CD8-/- and double KO mice at most time points. CD8-/- mice and the control group had near to superimposed kinetics. These results confirm the relevance of the present model.

Diagnosis of endometritis in the bitch: a new approach.

There are a few investigations into endometritis in the bitch and its relationship with failure to conceive remains unclear. This may be because of the difficulty in collecting uterine samples for further investigations. Recently, transcervical catheterization by vaginal endoscopy has been introduced allowing the evaluation of the endometrium. In this study, uterine cytology and bacteriology were evaluated in 26 infertile bitches. Endometritis was bacterial in origin in most cases (70% of affected bitches), but these results may be underestimated, as some other pathogens (anaerobic bacteria, mycoplasms and fungi) were not investigated. Endometritis, in our opinion, should be investigated in each case of unexplained infertility in bitches. The method used here seems reliable although defining more accurate classification criteria will improve the efficiency of this non-invasive technique.

Bartonella endocarditis: a pathology shared by animal reservoirs and patients.

Bartonellae were first recognized to cause endocarditis in humans in 1993 when cases caused by Bartonella quintana, B. elizabethae, and B. henselae were reported. Since the first isolation of Bartonella vinsonii subspecies berkhoffii from a dog with endocarditis, this organism has emerged as an important pathogen in dogs and an emerging pathogen in people. Subsequently, four types of B. vinsonii subsp. berkhoffii have been described, all of which have been associated with endocarditis in dogs. A limited number of dog endocarditis cases have also been associated with B. clarridgeiae, B. washoensis, B. quintana, and B. rochalimae. The second canine B. clarridgeiae endocarditis case is presented. The clinical and pathological characteristics of Bartonella endocarditis in dogs are similar to disease observed in humans, more often affecting the aortic valve, presenting with highly vegetative lesions with accompanying calcification, and in most instances high antibody titers. Pathological features in dogs include a combination of fibrosis, mineralization, endothelial proliferation, and neovascularization with variable inflammation. Endocarditis has also been described in animal species, which are the natural reservoir of specific Bartonella species, once thought to be solely healthy carriers of these pathogens. A few Bartonella endocarditis cases, including B. henselae, have been reported in cats in the USA and Australia. The second case of B. henselae type Houston I identified in the USA is presented. Furthermore, two cases of B. bovis endocarditis were recently described in adult cows from France. Finally, on-going investigation of valvular endocarditis in free-ranging Alaskan sea otters suggests the involvement of Bartonella species.

Inclusion of marbofloxacin in PMMA orthopaedic cement: an in vitro experimental study.

This in vitro experimental study investigated the feasibility for marbofloxacin, a veterinary fluoroquinolone antibiotic, to retain its antibacterial activity after its inclusion in polymethylmethacrylate (PMMA) cement. The assays were conducted on gelose cultures of various types of bacteria isolated from canine spontaneous osteomyelitis. The efficiency of the antibiotics was assessed by using an antibiogram method. Resistance of marbofloxacin to the temperature observed during PMMA polymerization, antimicrobial effect of galenic, useful concentrations and comparison with gentamicin (reference antibiotic for this use) were evaluated. Marbofloxacin retained its antimicrobial activity after being heated to high temperatures reached during polymerization. The more effective galenic form to incorporate into the PMMA monomer was the marbofloxacin powder and the appropriate concentration was 1/40 degrees . In this experiment, marbofloxacin included in PMMA showed a good antimicrobial activity; however this activity was lower than gentamicin added to PMMA on Gram + and Pseudomonas bacteria.Therefore, it seems useful to incorporate marbofloxacin to PMMA cement to treat, or to prevent, osteomyelitis associated with marbofloxacin sensitive bacteria. Nevertheless, the development of a marbofloxacin-PMMA cement requires further evaluation, especially pharmacological, biomechanical and clinical.

Effects of cow age and pregnancy on Bartonella infection in a herd of dairy cattle.

Bartonella spp. are small hemotropic bacteria infecting mammals. Four Bartonella species have been recently described in cattle and wild ruminants. To date, the biology and possible pathogenic role of Bartonella species isolated from ruminants are poorly understood. Therefore, a dairy herd of 448 cows and heifers was surveyed in order to establish the prevalence of Bartonella bovis and B. chomelii infections, the level of bacteremia, and the relationship between bacteremia and age or pregnancy status. The putative impact of Bartonella infection on production performance (individual milk cell count, milk yield) and reproductive status (success of artificial insemination [AI], placental retention, embryonic death, and abortion) was also assessed. The overall mean prevalence of B. bovis bacteremia was 59%, with the highest prevalence in heifers (92.5%). No B. chomelii was isolated, and 95% (114/120) of the B. bovis strains isolated and tested by PCR-restriction fragment length polymorphism belonged to type I. The level of bacteremia was higher in pregnant cows than in nonpregnant cows (P = 0.05), and the level of bacteremia rose during the last two-thirds of gestation (P < 0.001). There was no correlation between bacteremia and milk yield, individual milk cell count, success of first AI, interval between two calvings, or incidence of abortion and embryonic death. The interval from calving to first AI was shorter and the incidence of placental retention was lower in bacteremic animals than in nonbacteremic ones (P = 0.03 and P = 0.01, respectively).

First isolation of Mycobacterium microti (Llama-type) from a dog.

We report the first isolation of Mycobacterium microti from a dog with lesions of acute peritonitis. The isolate was demonstrated to be M. microti of Llama-Type by spoligotyping. Epidemiological implications of the isolation of this possibly zoonotic agent from a dog are discussed.