Microscopic detection of Leptospira bacterial organisms in urine sediment from a young dog with leptospirosis and a review of the pathobiology and diagnosis of canine leptospirosis

Samples collected from an 11-month-old Dachshund-mix dog with a history of acute azotemia, fever, and enlarged and irregular kidneys were received at the Colorado State Veterinary Diagnostic Laboratory (CSU VDL). The submitting veterinarians were concerned about lymphoma versus acute nephritis/pyelonephritis. The CSU clinical pathology laboratory received urine for urinalysis and kidney aspirates for cytologic evaluation. Urine had also been submitted for aerobic culture and Leptospirosis PCR, and serum was submitted for Lepto-5 microscopic agglutination testing (MAT). Upon examination of a wet mount of the urine sediment, technical staff noted “vibrating” clumps of granular-appearing material throughout the slide, which prompted the preparation of a stained sediment slide for pathologist review. Very small, faintly staining organisms were observed, and an attempt was made to picture-match these with published reports of Leptospira in dog urine, but none could be found. In addition, some references claimed that Leptospira organisms are not seen in urine with light microscopy. The suspicion that these organisms were Leptospira sp. was supported by the MAT results and later confirmed by PCR. The organisms subsequently exhibited strong positive immunolabeling for the Leptospira antigen. This case report provides a searchable record of Leptospira organisms visualized by routine light microscopy in dog urine during natural infection and a review of canine leptospirosis pathobiology and diagnosis.     

Human tularaemia associated with exposure to domestic dogs—United States, 2006–2016

Dogs have been implicated in the zoonotic transmission of numerous pathogens. Whereas cats are known to transmit Francisella tularensis to humans via bite and other routes, the role of dogs in facilitating infection is much less understood. We reviewed tularaemia case investigation records collected through national surveillance during 2006–2016 to summarize those with dog involvement, characterize the nature of dog‐related exposure and describe associated clinical characteristics. Among 1,814 human tularaemia cases, 735 (41%) supplemental case investigation records were available for review; and of those, 24 (3.3%) were classified as dog‐related. Median age of patients was 51 years (range: 1–82); 54% were female. Two thirds (67%) of cases presented with ulceroglandular/glandular tularaemia; pneumonic (13%) and oropharyngeal (13%) illness occurred less frequently. Dog‐related exposures were classified as follows: direct contact via bite, scratch or face snuggling/licking (n = 12; 50%); direct contact with dead animals retrieved by domestic dogs (n = 8; 33%); and contact with infected ticks acquired from domestic dogs (n = 4; 17%). Prevention of dog‐related tularaemia necessitates enhanced tularaemia awareness and tick avoidance among pet owners, veterinarians, health care providers and the general public.     

Early Detection of Brucella Canis via Quantitative Polymerase Chain Reaction Analysis

Canine brucellosis is a reportable zoonotic disease that can lead to canine reproductive losses and human infection through contact with infected urine or other genitourinary secretions. Although many locations require testing and euthanasia of positive dogs, current diagnosis is limited by the time required for seroconversion, for example, presence of B. canis‐specific antibodies. The goal of this study was to determine the diagnostic ability of Brucella canis‐specific quantitative polymerase chain reaction (qPCR) assay to detect B. canis in field samples prior to serological positivity for faster diagnosis and prevention of transmission within kennels or in households. Two kennels, one of which was located in the owner's home, were sampled following observation of suggestive clinical signs and positive serology of at least one dog. Specimens obtained were comparatively analysed via serology and qPCR analysis. 107 dogs were analysed for B. canis infection via qPCR: 105 via whole‐blood samples, 65 via vaginal swab, six via urine and seven via genitourinary tract tissue taken at necropsy. Forty‐five dogs were found to be infected with canine brucellosis via qPCR, of which 22 (48.89%) were seropositive. A statistically significant number (P = 0.0228) of qPCR‐positive dogs, 5/25 (20.00%), seroconverted within a 30‐day interval after initial serologic testing. As compared to serology, qPCR analysis of DNA from vaginal swabs had a sensitivity of 92.31% and specificity of 51.92%, and qPCR analysis of DNA from whole‐blood samples had a sensitivity of 16.67% and specificity of 100%. B. canis outer membrane protein 25 DNA qPCR from non‐invasive vaginal swab and urine samples provided early detection of B. canis infection in dogs prior to detection of antibodies. This assay provides a critical tool to decrease zoonotic spread of canine brucellosis, its associated clinical presentation(s), and emotional and economic repercussions.     

Generalized calcinosis cutis associated with disseminated paecilomycosis in a dog

A 4‐year‐old spayed female mixed breed dog was referred to the Michigan State University, Veterinary Teaching Hospital (MSU‐VTH) with vomiting, lethargy and anorexia of 2 weeks duration. Abdominal radiographs and ultrasonography showed hepatosplenomegaly. Cytological evaluation of ultrasound‐guided fine needle aspirates of the liver and spleen revealed fungal organisms and pyogranulomatous inflammation; fungal culture documented Paecilomyces variotii infection. The dog received antifungal therapy and supportive care. Multiple firm plaque‐like skin lesions, predominantly involving the inguinal region, developed 18 days after initial presentation and were diagnosed histopathologically as calcinosis cutis. While generalized calcinosis cutis has been reported in three dogs with blastomycosis and one dog with leptospirosis, the association with disseminated Paecilomyces spp. infection is novel.     

Epidemiological investigation of Leptospira spp. in a dairy farming enterprise after the occurrence of three human leptospirosis cases

An epidemiological investigation was conducted in an unvaccinated dairy farming enterprise in which three workers on one of the milking herds (Herd 1) were diagnosed with leptospirosis due to serovars Hardjo (H) (n = 2) and Pomona (P) (n = 1) between January and March 2015. Blood and urine samples were collected from milking cows in Herd 1 (N = 230) and Herd 2 (N = 400), rising one‐ (R1, N = 125) and rising two‐year‐old (R2, N = 130) replacement heifers, and four pigs associated with Herd 1, in March 2015. Sera were tested using the MAT for serovars H, P, Copenhageni (C), Ballum (B) and Tarassovi (T), and urine samples were tested by qPCR. Seventy‐five per cent of 109 cows in Herd 1 and 36% of 121 in Herd 2 were seropositive (≥48), predominantly to H and P, and 23% of 74 cows in Herd 1 and 1% of 90 cows in Herd 2 were qPCR positive. Fifty‐five per cent of 42 R2 heifers were seropositive to T. No R1 and 17% of 42 R2 heifers were qPCR positive. Subsequently, all cattle were vaccinated for H and P, and Herds 1 and 2 were given amoxicillin. After the booster vaccination, 7% of 91 in Herd 1, 2% of 82 in Herd 2 and 11% of 38 R1 heifers (sampled as R2) were PCR positive. After the amoxicillin treatment, no cows in Herd 1 and 5% of 62 cows in Herd 2 were urine PCR positive. Calves and pigs were seropositive to H, P, C and B. Vaccination and antibiotic treatment appeared effective in reducing the risk of exposure of workers to vaccine serovars. However, evidence of non‐vaccine serovars indicated that workers likely remain at risk of exposure to Leptospira.     

Variability in results of the microscopic agglutination test in dogs with clinical leptospirosis and dogs vaccinated against leptospirosis

Background: The microscopic agglutination test (MAT) is commonly used for the diagnosis of canine leptospirosis. In dogs it is sometimes suggested that the serogroup with the highest MAT titer is the infecting serogroup; however, this is not true in humans with confirmed leptospirosis. We sought to investigate the value of MAT results in predicting the infecting serogroup by comparing results across several laboratories and within individual dogs over time. Objectives: To examine the variability in MAT results across different laboratories in dogs recently vaccinated against leptospirosis, and in dogs with clinical leptospirosis, and to investigate variability over time in MAT results in individual dogs with leptospirosis. Animals: Eighteen dogs from a research colony, 9 of which had been vaccinated against leptospirosis, and 17 dogs clinically diagnosed with leptospirosis. Methods: Serum samples were submitted to up to 5 veterinary diagnostic laboratories for MAT titers from each dog on at least 1 occasion. MAT results also were followed over time in 6 dogs diagnosed with leptospirosis. Results: MAT results were discordant across different laboratories in dogs recently vaccinated against leptospirosis and in dogs with clinical leptospirosis. MAT results varied over time in individual dogs with the disease. Conclusions and Clinical Importance: The MAT is a valuable test for the diagnosis of leptospirosis in dogs, but it is unlikely that test results can be used to predict the infecting serogroup. Laboratories offering the MAT should consider participation in a proficiency scheme.     

Kidney Injury Molecule-1 in the detection of early kidney injury in dogs with leptospirosis

Renal damage, a common feature in canine leptospirosis, ranges from a subclinical affection to kidney dysfunction and death. Chances of recovery can be improved by early intervention. However, traditional biomarkers (serum urea and creatinine) have limited relevance for precocity. Kidney Injury Molecule-1 (KIM-1) is a transmembrane protein upregulated in early stages of tubular injury. This study evaluated the use of urinary KIM-1 to detect early renal injury in naturally occurring canine leptospirosis. This exploratory research included 30 dogs divided into two groups: (1) dogs with leptospirosis (n = 25) and (2) healthy dogs (n = 5). Leptospira sp. infection was diagnosed through urine PCR and/or direct bacteriologic culture and/or serology (single MAT titters ≥800). Additionally, stage of infection was further characterized in acute and subacute phases based on the onset of clinical symptoms from 3 to 7 days. Urinary KIM-1 (uKIM-1) concentrations were measured in both groups with a commercial canine ELISA kit.uKIM-1 levels were statistically different (P < 0.01) between the studied groups, especially in non-azotemic dogs (P = 0.0042). The biomarker showed 88 % sensibility to diagnosis of kidney injury at> 1.49 ng/mL cut-off. Urine KIM-1 was negatively correlated with urine specific gravity (USG) but accompanied histopathological evidence of renal degeneration, necrosis and regeneration processes, extending information on kidney health. Measurement of KIM-1 in the urine of canine patients was able to detect naturally occurring acute and subacute leptospirosis accompanied by tubular injury in early non-azotemic infections.     

A microbiological and serological study of leptospirosis among pigs in the Morogoro municipality,Tanzania

Serological and microbiological studies on leptospirosis in pigs from Morogoro municipality, Tanzania were carried out between October 2007 and May 2008. Serum samples (n = 385) from apparently healthy pigs were tested by microscopic agglutination test for antibodies against live cultures of six known Leptospira interrogans serovars: Pomona, Icterohaemorrhagiae, Ballum, Tarassovi, Grippotyphosa and Hardjo. Significant positive titres were detected in 4.42% (17/385) of all the tested serum samples. Asceptically collected samples, urine (n = 236) and kidney tissues (n = 214), were cultured in enriched Fletcher’s and Ellinghausen McCullough–Johnson and Harris media and assessed, at weekly intervals for 24 weeks, for growth by dark-field microscopy. Two leptospiral organisms were isolated from the urine samples. There was a statistical association between seroposivity and location that the subjects reside in (P < 0.05), whereas it was not significantly associated with sex nor age (P > 0.05). The evidence of pig exposure to different serovars and the isolation of the leptospiral organisms confirm that the infection is present in pigs although with an overall low prevalence. Apart from its economic importance on to the pig industry, this disease is a potential zoonotic public health risk in Tanzania, especially because of the lack of studies on leptospirosis among persons who handle pigs and pork products.     

Spatial epidemiological approaches to inform leptospirosis surveillance and control: A systematic review and critical appraisal of methods

Leptospirosis is a global zoonotic disease that the transmission is driven by complex geographical and temporal variation in demographics, animal hosts and socioecological factors. This results in complex challenges for the identification of high‐risk areas. Spatial and temporal epidemiological tools could be used to support leptospirosis control programs, but the adequacy of its application has not been evaluated. We searched literature in six databases including PubMed, Web of Science, EMBASE, Scopus, SciELO and Zoological Record to systematically review and critically assess the use of spatial and temporal analytical tools for leptospirosis and to provide general framework for its application in future studies. We reviewed 115 articles published between 1930 and October 2018 from 41 different countries. Of these, 65 (56.52%) articles were on human leptospirosis, 39 (33.91%) on animal leptospirosis and 11 (9.5%) used data from both human and animal leptospirosis. Spatial analytical (n = 106) tools were used to describe the distribution of incidence/prevalence at various geographical scales (96.5%) and to explored spatial patterns to detect clustering and hot spots (33%). A total of 51 studies modelled the relationships of various variables on the risk of human (n = 31), animal (n = 17) and both human and animal infection (n = 3). Among those modelling studies, few studies had generated spatially structured models and predictive maps of human (n = 2/31) and animal leptospirosis (n = 1/17). In addition, nine studies applied time‐series analytical tools to predict leptospirosis incidence. Spatial and temporal analytical tools have been greatly utilized to improve our understanding on leptospirosis epidemiology. Yet the quality of the epidemiological data, the selection of covariates and spatial analytical techniques should be carefully considered in future studies to improve usefulness of evidence as tools to support leptospirosis control. A general framework for the application of spatial analytical tools for leptospirosis was proposed.     

Evidence for Unapparent Brucella canis Infections among Adults with Occupational Exposure to Dogs

Human serological assays designed to detect brucellosis will miss infections caused by Brucella canis, and low levels of periodic bacteremia limit diagnosis by blood culture. Recent B. canis outbreaks in dogs and concomitant illnesses in caretakers suggest that unapparent human infections may be occurring. With more than a quarter of a million persons in occupations involving dogs, and nearly 80 million dog owners in the United States, this pathogen is an under‐recognized human health threat. To investigate occupational exposure to B. canis, we adapted a commercial canine serological assay and present the first controlled seroepidemiological study of human B. canis infections in recent years. 306 adults with occupational exposure to dogs and 101 non‐matched, non‐canine‐exposed subjects were enrolled. Antibodies were detected using the canine D‐Tec^® CB rapid slide agglutination test (RSAT) kit with a secondary 2‐mercaptoethanol (ME)‐RSAT. Results were validated on a blinded subset of sera with an additional RSAT and indirect enzyme‐linked immunoassay at the National Administration of Laboratories and Health Institutes (ANLIS) in Argentina. Seroprevalence ranged from 10.8% (RSAT) to 3.6% (ME‐RSAT) among canine‐exposed subjects. Kennel employees were more likely to test RSAT seropositive compared with other canine exposures (OR = 2.7; 95% CI, 1.3–5.8); however, low seroprevalence limited meaningful occupational risk factor analyses. Two seropositive participants reported experiencing symptoms consistent with brucellosis and having exposure to B. canis‐infected dogs; however, temporality of symptom onset with reported exposure could not be determined. D‐Tec^® CB results had substantial agreement with ANLIS assays (Cohen's kappa = 0.60–0.68). These data add to a growing body of literature suggesting that people occupationally exposed to dogs may be at risk of unapparent B. canis infection. It seems prudent to consider B. canis as an occupational public health concern and encourage the development of serological assays to detect human B. canis infections.     

Shedding and Seroprevalence of Pathogenic Leptospira spp. in Sheep and Cattle at a New Zealand Abattoir

A cross‐sectional study was carried out on sheep and cattle slaughtered at a New Zealand abattoir from September to November 2010 to investigate the supplier‐specific shedding rate, renal carriage rate and seroprevalence of leptospires. In the 2008/2009 season, this abattoir experienced three human leptospirosis cases from 20 staff, of which two were hospitalized. Urine, kidney and blood samples were collected from carcasses of 399 sheep (six suppliers, 17 slaughter lines) and 146 cattle (three suppliers, 22 slaughter lines). The urine and kidney samples were tested by quantitative real‐time PCR (qPCR), while serum samples (from coagulated blood samples) were tested by microscopic agglutination test (MAT). In total, 27% (73/274; 95% CI: 18–37) of urine samples tested positive by qPCR. Species‐specific shedding rates (prevalence of positive urine qPCR) were 31% (95% CI: 17–48) for sheep and 21% (95% CI: 14–30) for cattle. For 545 kidney samples tested, 145 were qPCR positive (27%; 95% CI: 17–39). The average prevalence of kidney qPCR positivity was 29% (95% CI: 17–45) for sheep and 21% (95% CI: 15–28) for cattle. Three hundred and thirty of 542 sampled sheep and cattle had antibodies against Leptospira borgpetersenii serovar Hardjobovis (Hardjobovis) and/or Leptospira interrogans serovar Pomona (Pomona), based on reciprocal MAT titre ≥1 : 48 (overall seroprevalence of 61%; 95% CI: 48–73). Seroprevalence was 57% (95% CI: 40–72) for sheep and 73% (95% CI: 59–83) for cattle. Among the seropositive animals, 41% (70/170; 95% CI: 30–54) were shedding (tested positive by urine qPCR) and 42% (137/330; 95% CI: 30–54) had renal carriage (tested positive by kidney qPCR). Some risk management options for abattoirs or farms to prevent human leptospirosis infections include vaccination of maintenance hosts, the use of personal protective equipment, and the application of urine qPCR to detect shedding status of stock as surveillance and as an alert.     

Evaluating the contact rate between companion dogs during dog walking and the practices towards potential cases of rabies among dog owners in Japan

This study aimed to examine the habits of dog walking in Japan using an internet survey of insured dog owners. About 96.4% of the respondents (n = 1,151) reported that they would take their dogs for a walk and they most frequently walk their dogs once or twice a day (75.9%) for 30 min to 1 hr (83.1%). The probability of a companion dog having contact with other dogs during dog walking was estimated to be 0.83 (95% confidence interval [CI]: 0.81–0.85), and the associated daily contact rate was estimated using log‐normal distribution with a mean of 2.73 (95% CI: 2.42–3.11) and a standard deviation (SD) of 6.39 (95% CI: 5.18–7.84). Multiple linear regression revealed that the contact rate is mainly influenced by the social behaviour of the owner and to a lesser degree by his/her demographic characteristics including the area of residence, the breed size of dog and the age of the owner. In addition, ten Likert items measured on a 5‐point scale were designed to assess the practices towards potential cases of rabies among dog owners. The respondents (n = 972) achieved a mean score of 2.99 (out of a full score of 4) with a SD of 0.90 in responding to situations related to dog bite incidents and injury from stray cat scratches during dog walking. They achieved a higher score in responding to situations related to sighting a stray or wild animal during dog walking and situations related to non‐specific clinical signs of rabies and bite injuries from stray dogs or wild animals during dog walking with a mean of 3.70 (SD = 0.58) and 3.84 (SD = 0.34), respectively. The level of best practice was also proved to be significantly associated with the demographic characteristics of the dog owner.     

Immunohistochemical detection of IgM and IgG in lung tissue of dogs with leptospiral pulmonary haemorrhage syndrome (LPHS)

Leptospiral pulmonary haemorrhage syndrome (LPHS) is a severe form of leptospirosis. Pathogenic mechanisms are poorly understood. Lung tissues from 26 dogs with LPHS, 5 dogs with pulmonary haemorrhage due to other causes and 6 healthy lungs were labelled for IgG (n = 26), IgM (n = 25) and leptospiral antigens (n = 26). Three general staining patterns for IgG/IgM were observed in lungs of dogs with LPHS with most tissues showing more than one staining pattern: (1) alveolar septal wall staining, (2) staining favouring alveolar surfaces and (3) staining of intra-alveolar fluid. Healthy control lung showed no staining, whereas haemorrhagic lung from dogs not infected with Leptospira showed staining of intra-alveolar fluid and occasionally alveolar septa. Leptospiral antigens were not detected. We conclude that deposition of IgG/IgM is demonstrable in the majority of canine lungs with naturally occurring LPHS, similar to what has been described in other species. Our findings suggest involvement of the host humoral immunity in the pathogenesis of LPHS and provide further evidence to support the dog as a natural disease model for human LPHS.     

Strategies of the control of an outbreak of leptospiral infection in dairy cattle in Northeastern Brazil

The aim of the present study was to describe the strategies of the control of an outbreak of leptospiral infection in dairy cattle in Maranhão State, Northeastern Brazil. In the period from January to July 2015, 18 (17%) out of 106 cows presented abortion, six (5.7%) stillbirth, and 12 (11.3%) repeated estrus, totaling 24 animals with reproductive problems. The diagnosis of leptospirosis was based on serology (microscopic agglutination test—MAT), bacteriological culture, and polymerase chain reaction (PCR). Antibiotic therapy, vaccination protocols, and changes in management practices were suggested as control measures. Of all animals on the farm (n?=?280), 136 (48.6%) were seropositive for at least one serovar of Leptospira sp. No pure leptospiral culture was obtained. Eight of the animals with reproductive problems yielded positive PCR results (vaginal fluid of seven animals and urine and vaginal fluid of one animal). Genetic sequencing of a vaginal fluid/urine PCR-positive sample revealed Leptospira borgpetersenii. One year after the adoption of control measures, no reproductive problems were observed. Thus, leptospirosis probably caused the reproductive failures in the herd, and the control and prevention measures implemented were efficient in controlling the disease.

     

Detection of fluoroquinolone resistance level in clinical canine and feline Escherichia coli pathogens using rapid real-time PCR assay

Fluoroquinolones are used to treat infections caused by Escherichia coli in canine and feline veterinary patients, particularly those infecting the urinary tract. The gyrA gene is a primary target causing fluoroquinolone resistance in Gram negative coliforms, with mutations in codons 83 and 87 generally associated with high-level of resistance E. coli clinical isolates. We have developed a fluorescence resonance energy transfer (FRET) quantitative PCR to identify enrofloxacin-resistance in clinical E. coli isolates that carry mutations in codons 83 and 87 of gyrA. This real-time quantitative PCR assay is rapid, economical, and sensitive compared with cultured antimicrobial susceptibility testing. The assay identified as few as four genome copies per reaction from culture and 19 genome copies in urine. For the 70 isolates tested, the sensitivity was 87.5% (95% CI = 75–95.3%) (n = 42/48), specificity was 100% (95% CI = 87.3–100%) (n = 22/22), whereas accuracy was 91.4% (95% CI = 82.3–97%) (n = 64/70). Furthermore, we were able to accurately differentiate between the wild type and mutants E. coli directly from infected canine urine samples (n = 5) within 2 h. These results were confirmed by sequence alignments of the PCR products and comparison with the susceptibility testing. The FRET-PCR assay appears to have promising clinical application as an early diagnostic tool for rapid and sensitive detection and differentiation of the level of fluoroquinolone resistance among clinical E. coli isolates that may facilitate design of the dosing regimen.     

Outbreak of Campylobacteriosis Associated with Raccoon Contact at a Wildlife Rehabilitation Centre,Minnesota, 2013

Campylobacteriosis is an enteric illness caused by bacteria of the genus Campylobacter. There are approximately 900 culture‐confirmed cases of campylobacteriosis reported annually to the Minnesota Department of Health (MDH). Case patients are interviewed about risk factors, including foods eaten, recreational and drinking water exposures and animal contact. In September 2013, MDH identified two Campylobacter jejuni cases who reported working at the same wildlife rehabilitation centre before illness onset. This report describes the investigation, which used a case–control study design, and identified 16 additional ill persons, for a total of 18 ill persons. Both cases and controls reported working with a variety of animals, including squirrels, chipmunks, mice, raccoons, opossums, rabbits, songbirds, waterfowl and reptiles. In univariate analyses, contact with a number of different animal species was significantly associated with illness, including raccoons (odds ratio [OR], 11.1; P < 0.001), chipmunks (OR, 3.65; P = 0.01), opossums (OR, 4.38; P = 0.005), mice (OR, 4.18; P = 0.01) and rabbits (OR, 4.36; P = 0.003). In a multivariate model, contact with raccoons was the only exposure independently associated with illness (adjusted OR, 12.2; P = 0.01). Bacterial culture and subtyping of the outbreak strain of C. jejuni from raccoon faecal samples further implicated raccoons as the source of the outbreak. Not all of the cases reported handling raccoons, suggesting that environmental contamination contributed to transmission. MDH worked with the wildlife rehabilitation centre's management to strengthen biosecurity and infection control protocols.     

Human Brucella canis Infection and Subsequent Laboratory Exposures Associated with a Puppy,New York City, 2012

Human Brucella canis infection incidence is unknown. Most identified cases are associated with pet dogs. Laboratory‐acquired infections can occur following contact with Brucella spp. We identified a paediatric B. canis case, the source and other exposed persons. A 3‐year‐old New York City child with fever and dyspnoea was hospitalized for 48 h for bronchiolitis. After her admission, blood culture grew B. canis, she was prescribed anti‐microbials and recovered. B. canis was also isolated from blood of the child's pet dog; these isolates were genetically similar. The dog originated from an Iowa breeding facility which was quarantined after identification of the dog's infection. Additionally, 31 laboratory workers were exposed and subsequently monitored for symptoms; 15 completed post‐exposure prophylaxis. To our knowledge, this is the first report strongly suggesting B. canis zoonotic transmission to a child in the United States, and highlights the need for coordinated control policies to minimize human illness.     

Monitoring of leptospirosis seroprevalence in a colony of captive collared peccaries (Tayassu tajacu) from the Peruvian Amazon

Leptospirosis, an endemic zoonoses, is maintained in the environment by several wildlife species in the Peruvian Amazon. In order to evaluate the possible role of collared peccaries (CP) in the maintenance this disease, two serological surveys of leptospirosis were performed and zootechnical parameters were monitored in a captive CP colony in an interval of 27 months. Total seroprevalence changed from 100% (n = 27) to 86.4% (n = 22), with reactions to a diversity of serogroups of zoonotic importance. Serological reactions to Leptospira licerasiae serogroup Iquitos, a new species recently identified locally and Leptospira interrogans serogroup Icterohaemorrhagiae were highly prevalent. The observation of leptospiral antibodies in both surveys, changes on serological reactions to different serogroups in large part of the herd and poor reproductive performances, provided an indication of the role of CP farms as a favourable environment for maintaining leptospirosis. Further research regarding the role of CP in the epidemiology of leptospirosis in the Peruvian Amazon is encouraged.     

Proof-of-concept study: Evaluation of plasma and urinary electrolytes as markers of response to L-asparaginase therapy in dogs with high-grade lymphoma

Response to chemotherapy is one of the most important prognostic factors in dogs with lymphoma. The objective of this feasibility study was to evaluate if clinical responses to a specific cytotoxic agent (L-asparaginase) could be anticipated by measuring analyte concentrations in plasma and urine concentrations of lymphoma-bearing dogs. We hypothesized that potassium and phosphate concentrations in plasma and urine would be higher in dogs that completely responded to therapy. Plasma and urine samples of dogs with lymphoma were obtained before 12 and 24 hours after intramuscular L-asparaginase injections. Peripheral lymph node volumes were evaluated according to the Veterinary Cooperative Oncology Group standardized criteria. Plasma and urine electrolyte, calcium, phosphate, creatinine, urea, total protein, and albumin concentrations were measured, and the fractional excretions of each electrolyte were calculated. Statistical analyses compared complete vs partial responders using a linear regression model. Contrast analyses were also performed to differentiate the mean of each group, with adjustments made with the Benjamini-Hochberg procedure. Fourteen dogs were included, eight with complete responses, and six with partial responses. Plasma phosphate concentrations were significantly higher at 12 hours (P = .0003) and 24 hours (P = .009) after complete responses to therapy. This study demonstrates the potential use of plasma and urine analyte monitoring after chemotherapy induction. Plasma phosphate measurements represent a potential indicator of early responses to L-asparaginase therapy. Larger population studies are warranted to confirm these preliminary results.     

Development of an indirect enzyme-linked immunosorbent assay for the detection of leptospiral antibodies in dogs.

Serology plays an important role in the diagnosis of leptospirosis. Few laboratories have the resources, expertise, or facilities to perform the microscopic agglutination test (MAT). Thus, there is a need for a rapid and simple serological test that could be used in any diagnostic laboratory. In this study, a genus-specific, heat-stable antigenic preparation from Leptospira interrogans serovar pomona was used in an enzyme-linked immunosorbent assay (ELISA) for the detection of leptospiral antibodies in dog sera. This antigenic preparation reacted with rabbit antisera against L. interrogans serovars bratislava, autumnalis, icterohaemorrhagiae and pomona and with rabbit antiserum against L. kirschneri serovar grippotyphosa. The ELISA showed a relative specificity of 95.6% with 158 dog sera which were negative at a dilution of 1:100 in the MAT for serovars pomona, bratislava, icterohaemorrhagiae, autumnalis, hardjo, and grippotyphosa. The relative sensitivity of this assay with 21 dog sera that revealed serovars MAT titres of > or =100 to different serovars was 100%. This assay is easily standardized, technically more advantageous than MAT, and uses an antigenic preparation that can be routinely prepared in large amounts. It was concluded that this ELISA is sufficiently sensitive test to be used as an initial screening test for the detection of leptospiral antibodies in canine sera, with subsequent confirmation of positive test results with the MAT.