Diagnosis and prevalence of leptospira infection in aborted and stillborn horses

A study was conducted to evaluate a recently available fluorescent antibody test (FAT) conjugate for the detection of leptospires in tissues of aborted and stillborn horses, to determine the leptospira antibody titers and compare serologic test results with FAT results, and to determine the prevalence of leptospira-induced abortions and stillbirths in the equine population of central Kentucky. From July 1, 1988 through June 30, 1989, 15 (2.5%) of 594 submissions (fetuses, stillborn foals, and/or placentas) were diagnosed as leptospirosis by the FAT (14 of 15 tested) and/or microscopic agglutination test (12 of 14 tested). Of the 12 serologically positive fetal fluids, 10 had high tigers against Leptospira interrogans serovar pomona and 2 against serovar grippotyphosa.     

Clustering of reported cases of leptospirosis among dogs in the United States and Canada

A retrospective study was conducted to determine whether cases of leptospirosis diagnosed among dogs in the United States and Canada were clustered in time, space, or time-and-space. The study population was estimated--based on first examination of each dog--to consist of 1,035,366 dogs examined at 22 veterinary teaching hospitals between 1983 and 1998. The veterinary medicine database was searched for records of dogs in which a diagnosis of leptospirosis was made during the study period. The scan statistic (Poisson model) was used to examine for clustering of cases, using recorded date of diagnosis and the location of the veterinary teaching hospital at which the diagnosis was made. Three-hundred and forty dogs diagnosed with leptospirosis were identified. Cases were clustered in the period of 1992-1998. A cluster of dogs diagnosed at three veterinary teaching hospitals located in the midwest of the United States was identified. The composition of this spatial cluster was influenced by the age and gender distribution of the population at-risk. A cluster of cases involving 143 dogs diagnosed with leptospirosis between 9 April 1993 and 14 December 1998 at six veterinary teaching hospitals located in the midwest of the United States was identified. This time-space cluster was not influenced by the age, gender or breed distribution of the population of dogs presented at veterinary teaching hospitals during the study period. Leptospirosis was clustered during the 1990s in the midwest of the United States. Dog risk factors (age, gender, breed) do not explain the clustering that was observed. Leptospira serovar or environmental factors might be responsible for the clustering of canine leptospirosis in the United States and Canada.     

Seasonality of canine leptospirosis in the United States and Canada and its association with rainfall

A retrospective study was undertaken to describe seasonal patterns of leptospirosis diagnosed at veterinary teaching hospitals in the United States and Canada, and to determine if occurrence of cases (diagnoses) was associated with rainfall. The veterinary medicine database (VMDB) was searched for records of dogs in which a diagnosis of leptospirosis was made, and average monthly rainfall recorded at the city where each hospital is located was calculated from historical data. Time-series analysis was used to identify the variables that best described occurrence of cases of leptospirosis. Three-hundred and forty dogs were diagnosed with leptospirosis at 22 veterinary teaching hospitals between 1983 and 1998. Most cases (n = 184) were diagnosed between August and November during each year of the study. A significant correlation (r = 0.41) was detected between the number of cases of leptospirosis diagnosed and average rainfall recorded 3-months prior to diagnosis. The best-fitting (Akaike's corrected information criterion = 2.01) regression on the leptospirosis case series included cases diagnosed in the previous month and 12-months previously, and rainfall recorded 3 months previously. Leptospirosis has a seasonal distribution (late summer to fall), and rainfall can be used to predict the occurrence of leptospirosis. Leptospirosis can affect the health of dogs seriously and its prevalence is increasing, but the disease is preventable by vaccination.     

Recovery of Arcobacter spp. from nonlivestock species

The genus Arcobacter encompasses campylobacter-like organisms that grow in air at 25 degrees C. Arcobacter has been detected or isolated from clinically healthy livestock as well as aborted fetuses and has been presumptively identified as either Campylobacter or Leptospira, based on its growth in selective semisolid media. Because reports from nonlivestock species are limited, this study examined nine presumptive isolates of Arcobacter spp. from an alpaca (Vicugna pacos), black rhinoceros (Diceros bicornis), white rhinoceros (Ceratotherium simum), gorilla (Troglodytes gorilla), gazelle (Eudorcas thomsoni), rhea (Rhea americana), and aborted equine fetuses. Seven of these nine phenotypically identified isolates of Arcobacter were confirmed by a multiplex polymerase chain reaction assay. The remaining two isolates were subsequently identified as Arcobacter skirrowii (Case 5) and Campylobacter jejuni (Case 6) by sequence analysis of a 527-base pair fragment of the 16S rRNA gene. Together, these cases underscore the challenges to a clinical laboratory of identifying Arcobacter in cases which mimic vibrionic abortion or leptospirosis.     

Prevalence and serovars of leptospira involved in equine abortions in central Kentucky during the 1990 foaling season.

A study to determine the prevalence of leptospira-induced abortions in the central Kentucky equine population during the 1990 foaling season and to determine the leptospira serovars responsible was conducted. From July 1, 1989 through June 30, 1990, 32 (4.4%) of 726 submissions (fetuses, stillborn foals, and/or placentas) were diagnosed as leptospirosis by the fluorescent antibody test and/or microscopic agglutination test. Attempts were made to isolate leptospires from the fetal tissues and/or the dam's urine in 31 of these cases. Leptospira interrogans serovar kennewicki was isolated from 11 (35.5%) and serovar grippotyphosa from 2 (6.5%) of the 31 cases. Of 12 cases that were culture negative with serologically positive fetal fluids, 8 had titers against serovar pomona, 1 against bratislava, 1 against grippotyphosa, 1 against hardjo, and 1 against both bratislava and pomona.     

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.     

An old disease with a new face: canine leptospirosis does not lose its relevance

The clinical features of the disease are presented based on retrospective analysis of the records of eleven dogs diagnosed with leptospirosis using clinical signs and results of the microagglutination test (MAT) between 1991 and 1996. Additionally, Leptospira titres were determined in 30 healthy dogs and 20 hospitalised dogs without clinical or laboratory evidence of leptospirosis. A positive titre for L. grippotyphosa, L. pomona, L. bratislava, L. australis, L. icterohaemorrhagiae and/or L. canicola was found in 16 normal dogs and only one hospitalised patient. Eight of these dogs had titres of > or = 1:800. Only one of them had been vaccinated shortly before sampling. These results suggest that many dogs from the surroundings of Bern, Switzerland have contact with various Leptospira interrogans serovars. In ten healthy dogs, the Leptospira titre was determined before and four weeks after vaccination with leptospiral antigen. Only two of the dogs showed a serologically measurable response to the antigen contained in the vaccine. In dogs MAT titers presumably do not reliably reflect the immune status against leptospiral infections.     

RADIOGRAPHIC PROTOCOL AND NORMAL ANATOMY OF THE HIND FEET IN THE WHITE RHINOCEROS (CERATOTHERIUM SIMUM)

Foot pathology is a common and important health concern in captive rhinoceroses worldwide, but osteopathologies are rarely diagnosed, partly because of a lack of radiographic protocols. Here, we aimed to develop the first radiographic protocol for rhinoceros feet and describe the radiographic anatomy of the white rhinoceros (Ceratotherium simum) hind foot (pes). Computed tomographic images were obtained of nine cadaver pedes from seven different white rhinoceroses and assessed for pathology. A single foot deemed free of pathology was radiographed using a range of different projections and exposures to determine the best protocol. 3D models were produced from the CT images and were displayed with the real radiographs to describe the normal radiographic anatomy of the white rhinoceros pes. An optimal radiographic projection was determined for each bone in the rhinoceros pes focusing on highlighting areas where pathology has been previously described. The projections deemed to be most useful were D60Pr‐PlDiO (digit III), D45Pr45M‐PlDiLO (digit II), and D40Pr35L‐PlDiLO (digit IV). The primary beam was centered 5–7 cm proximal to the cuticle on the digit of interest. Articular surfaces, ridges, grooves, tubercles, processes and fossae were identified. The radiographic protocol we have developed along with the normal radiographic anatomy we have described will allow for more accessible and effective diagnosis of white rhinoceros foot osteopathologies.     

Neosporosis in a white rhinoceros (Ceratotherium simum) calf

A 16-day-old white rhinoceros calf died suddenly while in excellent condition and showing no obvious previous clinical signs. It was the 9th calf of a mature female kept free-ranging with 11 other rhinoceros as well as various other game species on a 2000 hectare game breeding centre adjacent to the town of Lichtenburg and outlying cattle farmlands in the Northern Province. At post mortem examination, death was ascribed to heart failure. There was marked multifocal to coalescing subacute parasitic myocarditis with numerous protozoan bradyzoite cysts and free tachyzoites present amongst the predominantly round cell inflammatory infiltrate. The coccidian was positively identified as Neospora sp. using both polyclonal and murine monoclonal Neospora caninum antibody immunohistochemistry in the avidin-biotin technique. The parasites stained poorly with Toxoplasma gondii-specific immunoperoxidase staining. Ultrastructurally, a section of a bradyzoite-containing cyst, as well as tachyzoites, were largely but not totally consistent with those described for Neospora caninum. The dam showed no sign of illness. Neosporosis affecting white rhinoceros (Ceratotherium simum) has not previously been reported. A summarised overview of neosporosis from selected publications and a recent review is given.     

Prevalence of antibodies to Leptospira interrogans serovar hardjoin bulk tank milk from unvaccinated irish dairy herds

: Bulk tank milk samples, collected from 347 herds throughout the Republic of Ireland using a sampling frame based on seven milk-recording organisations, were tested by ELISA for antibodies to Leptospira interrogans serovar hardjo. These herds, which had not been vaccinated against leptospirosis within the previous five years, were categorised according to their province, milk-recording organisation and size. Two-hundred-and-seventy-three herds (79%) had a positive ELISA titre. Both the probability of a herd being seropositive and the antibody level in the herd milk sample were affected by the province (P < 0.05 and P < 0.01, respectively) and the herd size category (P < 0.05 and P < 0.01, respectively). Larger herds were significantly more likely to have positive reactions and higher mean concentrations of antibody. It was concluded that a high proportion of unvaccinated Irish dairy herds have been exposed to infection with Leptospira hardjo.     

Acute lymphoblastic leukemia in a juvenile southern black rhinoceros (Diceros bicornis minor).

A 21-mo-old female southern black rhinoceros (Diceros bicornis minor) developed acute upper respiratory dyspnea in association with lymphadenopathy and marked immature lymphocytosis. A diagnosis of acute lymphoblastic leukemia was reached on the basis of the morphologic and cytochemical characteristics of peripheral lymphoblasts. Antineoplastic chemotherapy included administration of cytarabine, cyclophosphamide, vincristine, and doxorubicin, with clinical remission achieved 19 days after initiation of treatment. The rhinoceros died, however, of congestive heart failure, presumably secondary to doxorubicin cardiotoxicity and a particular sensitivity of rhinoceros myocardial tissue to free hydroxyl radicals. The pharmacologic effects of any therapeutic agent need to be carefully considered before use in the black rhinoceros, especially within the context of the unique physiology of this species.     

Idiopathic hemorrhagic vasculopathy syndrome in seven black rhinoceros

Idiopathic hemorrhagic vasculopathy syndrome (IHVS) was diagnosed in 7 black rhinoceros; this newly described syndrome is characterized by severe body swelling in conjunction with a rapid and profound decrease in Hct. The disorder may be acute or chronic, may recur, and is potentially fatal. Five of the rhinoceros survived an initial episode of IHVS, and 2 of these 5 survived a recurrent episode of IHVS. Two rhinoceros died during treatment of IHVS. Treatment protocols varied, but all 7 rhinoceros received broad-spectrum antibiotics, because an infectious cause was suspected. All rhinoceros also received nonsteroidal antiinflammatory drugs and supportive care. Idiopathic hemorrhagic vasculopathy syndrome has many similarities to other vasculopathies of domestic animals, such as equine purpura hemorrhagica, but it also appears to have unique identifying features. It has been hypothesized that IHVS may be an immune response to an as yet unidentified infectious agent. Thorough and extensive testing has not identified the potential causative agent, nor the factors that predispose some black rhinoceros to developing IHVS. Further research into the rhinoceros immune system is ongoing and should help elucidate the mechanisms through which IHVS develops.     

Gross anatomy and ultrasonographic images of the reproductive system of the Sumatran rhinoceros (Dicerorhinus sumatrensis)

The Sumatran rhinoceros (Dicerorhinus sumatrensis) is the smallest of all the rhino species. It is one of the rarest mammals in the world and is in imminent danger of extinction. A study was carried out on seven wild-caught females, three wild-caught males and one captive born female Sumatran rhinoceros at the Sumatran Rhinoceros Breeding Centre in Sungai Dusun, Selangor, Malaysia, beginning 1990. As a result of the paucity of scientific information on the reproductive biology of the Sumatran rhinoceros, this study was conducted to obtain information, which could assist in the captive breeding of this endangered and near extinct species. The anatomy of the reproductive system was based on two post-mortem specimens and transrectal real-time ultrasonography in six adult females. Genitalia of the Sumatran rhinoceros were similar to those of other species of rhinoceroses. The cervix consisted of several folds, the uterus was bicornuate with a short body and prominent horns and the ovaries were completely covered by the fimbriated end of the fallopian tube. The internal genitalia could be imaged by ultrasonography. The testes were located within a pendulous scrotum. Two lateral projections were located at the base of the penis. A well-defined process glandis was present at the tip of the penis. The accessory sex glands and the testes could be imaged by ultrasonography.     

Clinical application of a polymerase chain reaction assay for diagnosis of leptospirosis in dogs

OBJECTIVE: To evaluate the use of a polymerase chain reaction (PCR) assay on urine samples for diagnosis of leptospirosis in dogs. DESIGN: Prospective case study. ANIMALS: 132 dogs with clinical signs suggestive of leptospirosis and 13 healthy dogs. PROCEDURE: PCR testing was performed on urine samples to detect leptospiral DNA; results were compared with results of conventional criteria for the diagnosis of leptospirosis. RESULTS: Leptospirosis was diagnosed in 8 dogs via established criteria; all these dogs had positive results of PCR assay, including 1 dog with positive results before seroconversion developed. A positive PCR assay result was also obtained in 16 dogs that did not have a confirmed diagnosis of leptospirosis. In the 8 dogs that had a confirmed diagnosis of leptospirosis, serovars pomona (n = 3 dogs), grippotyphosa (2), canicola (2), and bratislava (1) were identified serologically. The remaining 121 dogs all had a diagnosis other than leptospirosis or were healthy. For PCR testing on urine, sensitivity was 100%, specificity was 88.3%, positive predictive value was 33%, and negative predictive value was 100%. CONCLUSIONS AND CLINICAL RELEVANCE: Positive PCR test results prior to seroconversion may have value in establishing an early diagnosis. Positive results in dogs that had signs consistent with leptospirosis despite failing to meet established criteria for leptospirosis raise questions regarding the sensitivity of serologic testing in diagnosis of leptospirosis. Serovars pomona, grippotyphosa, and canicola were most common.     

Ehrlichia canis-related polyarthritis in a dog

Ehrlichia canis-related polyarthritis was diagnosed in a 7-month-old Boxer. The diagnosis was based on intraneutrophilic morulae found in synovial fluid specimens, thrombocytopenia, a positive result for indirect fluorescent antibody testing for E canis, the presence on the dog of the known vector of E canis infection (Rhipicephalus sanguineus), and a favorable response to treatment with tetracycline hydrochloride. The dog has had no recurrence of lameness for 18 months after cessation of treatment.     

Pulmonary Abnormalities in Dogs with Leptospirosis

Background: Leptospirosis in dogs is a multiorgan disease affecting mostly kidneys and liver. Objectives: The objective was to characterize prevalence, clinical, and radiological features and outcome of dogs with leptospirosis and pulmonary abnormalities. Animals: Fifty dogs with leptospirosis. Methods: Medical records of dogs diagnosed with leptospirosis at the Small Animal Clinic, Berlin, were reviewed. Diagnosis was based on microscopic agglutination test, blood or urine polymerase chain reaction, and histopathology. Based on clinical and/or radiological signs, patients were grouped into dogs with lung abnormalities (group 1) or without (group 2). Severity of respiratory distress was scored as mild to moderate (grade 1) or severe (grade 2). Thoracic radiographs were scored based on pulmonary changes and location as grade 1 (caudal interstitial pattern), 2 (generalized mild to moderate reticulonodular interstitial pattern), or 3 (generalized severe reticulonodular interstitial pattern with patchy alveolar consolidations). Results of CBC and biochemistry were compared between groups. Results: Thirty‐five dogs had radiological pulmonary changes (grade 1: 5; grade 2: 14; grade 3: 16); 31 of them had pulmonary distress (grade 1: 13, grade 2: 18). Sixty‐seven percent of the dogs with dyspnea grade 2 were mainly euthanized because of respiratory distress. Fifteen percent of the dogs with dyspnea grade 1 and 21% without clinical respiratory signs were euthanized because of acute renal failure or sepsis. Conclusions and Clinical Importance: In 70% of dogs with leptospirosis pulmonary changes were detected. Lung involvement represented a severe complication causing increased case fatality depending on the severity of respiratory distress.     

青海地区猪流行性腹泻的病原诊断

对青海地区疑似猪流行性腹泻病料进行了细菌学和轮状病毒核酸电泳的诊断。试验结果:排除了致病菌和轮状病毒的病原感染;乳猪回归试验,复制出与自然病例相同的临床症状;病料经处理后与猪流行性腹泻病毒免疫血清发生特异性凝集;与猪传染性胃肠炎病毒荧光抗体染色呈阴性反应;与猪流行性腹泻病毒免疫血清进行间接荧光染色时,在细胞浆内见到了特异性荧光;电镜观察到冠状病毒科的病毒颗粒。从而确诊青海地区猪流行性腹泻病原为猪流行性腹泻病毒,代号为PEDV—青毒1株。     

A multipronged approach for the detection of leptospirosis in captive sloth bears (Melursus ursinus) in Agra and Bannerghatta sloth bear rescue centers in India

Leptospirosis is an exacerbating factor responsible for the drastic decline of sloth bear population in India. In this study, a multipronged approach based on antigen detection using Polymerase Chain Reaction (PCR) employing G1/G2 and LigBF/LigBR primers, antibody detection using Microscopic Agglutination Test (MAT) and recombinant LigBCon1-5 antigen based Latex Agglutination Test (rLigBCon1-5 LAT), serum biochemistry using hepatic (serum glutamate oxalo acetic transaminase (SGOT) and serum glutamate pyruvic transaminase (SGPT) and renal biomarkers (blood urea nitrogen (BUN) and Creatinine) and gross/histopathological evidence in liver and kidneys were employed to investigate leptospirosis in captive sloth bears. A total of 133 serum samples collected from Agra (n=113) and Bannerghatta (n=20) sloth bear rescue centers were screened using MAT and rLigBCon1-5 LAT. A total of 87 and 78 sera tested positive by MAT and LAT respectively. Pyrogenes was the leading serovar obtained using MAT followed by Icterohaemorrhagiae, Javanica, Grippotyphosa, Canicola and Tarassovi. The relative sensitivity, specificity and accuracy of rLigBCon1-5 LAT in comparison to MAT were 89.66%, 100% and 93.23% respectively. PCR performed on hepatic and renal tissues showed amplicon of 285 and 219 base pairs for G1/G2 and LigBF/LigBR primers respectively. Gross evidence (icteric liver, severely engorged hepatic sinusoids, congested kidneys with necrotic white spots on sub capsular surface), histopathology (severe hepatic degeneration and tubulointerstitial nephritis) and elevated hepatic/renal biomarkers were suggestive of leptospirosis. This study suggests that rLigBCon1-5 LAT can be employed as a pen-side test for detecting leptospirosis in sloth bears.     

Urinalysis in three species of captive rhinoceros (Rhinoceros unicornis, Dicerorhinus sumatrensis, and Diceros bicornis)

This study reports urinalysis values for three species of captive rhinoceros (Rhinoceros unicornis, Dicerorhinus sumatrensis, and Diceros bicornis) and evaluates individual and species differences. Repeated urinalysis was conducted on 11 individuals to establish normal reference ranges. Although no individual or species differences existed in urinary values for pH, all species differed in specific gravity. Rhinoceros urine demonstrated many physical and chemical properties similar to that of the horse, but reliability of this comparison was limited. Urinary pH in the rhinoceros was within range of that established for the horse and other large herbivores. However, all rhinoceros species exhibited urinary specific gravities below the lower limit of the normal equine reference range. Comparative urinalysis using an outside laboratory source confirmed the results of this study and illustrated the value of conducting in-house analysis. These results are the first data available on reference ranges for urine parameters in the greater one-horned, Sumatran, and African black rhinoceros and provide a useful diagnostic tool for the veterinary care of individuals in captivity.     

Black rhinoceros (Diceros bicornis) and domestic horse (Equus caballus) hindgut microflora demonstrate similar fermentation responses to grape seed extract supplementation in vitro

The domestic horse is used as a nutritional model for rhinoceros maintained under human care. The validity of this model for browsing rhinoceros has been questioned due to high prevalence of iron overload disorder (IOD) in captive black rhinoceros (Diceros bicornis), which is associated with high morbidity and mortality. Iron chelators, such as tannins, are under investigation as dietary supplements to ameliorate or prevent IOD in prone species. Polyphenolic compounds variably affect microbial fermentation, so the first objective of this experiment was to evaluate the effects of grape seed extract (GSE; a concentrated source of condensed tannins; CT) on black rhinoceros hindgut fermentation. Equine nutrition knowledge is used to assess supplements for rhinoceros; therefore, the second objective was to evaluate the domestic horse model for black rhinoceros fermentation and compare fermentation responses to GSE using a continuous single‐flow in vitro culture system. Two replicated continuous culture experiments were conducted using horse and black rhinoceros faeces as inoculum sources comparing four diets with increasing GSE inclusion (0.0%, 1.3%, 2.7% and 4.0% of diet dry matter). Diet and GSE polyphenolic compositions were determined, and sodium sulphite effect on neutral detergent fibre extraction of CT‐containing forages was tested. Increasing GSE inclusion stimulated microbial growth and fermentation, and proportionally increased diet CT concentration and iron‐binding capacity. Horse and black rhinoceros hindgut microflora nutrient digestibility and fermentation responses to GSE did not differ, and results supported equine fermentation as an adequate model for microbial fermentation in the black rhinoceros. Interpretation of these results is limited to hindgut fermentation and further research is needed to compare foregut digestibility and nutrient absorption between these two species. Supplementation of GSE in black rhinoceros diets up to 4% is unlikely to adversely affect hindgut nutrient digestibility or microbial viability and fermentation.