Carrier Animals Research Articles

Leptospira spp.in Wild Rodents, Canary Islands

<i>Leptospira spp.</i>in Wild Rodents, Canary Islands

Salmonella Enterica Serovar Typhimurium DT104 Invasion is Not Enhanced by Sub-Inhibitory Concentrations of the Antibiotic Florfenicol

The incidence of multi-drug resistant Salmonella has increased globally over the past several decades and has become a major public health concern. Isolates of Salmonella enterica serovar Typhimurium DT104 are resistant to five or more antibiotics, including florfenicol, and have been associated with enhanced virulence in livestock and humans. Because sub-inhibitory concentrations of some antibiotics have been found to modulate invasion of certain bacteria under specific conditions, the effect of florfenicol on S. Typhimurium DT104 invasion was evaluated. Three clinical bovine isolates were exposed to sub-inhibitory concentrations of florfenicol for 30 minutes to establish the initial response to the antibiotic. HEp-2 cellular invasion assays, as well as expression analyses of invasion-related genes, demonstrated that the invasiveness of the S. Typhimurium DT104 isolates was not enhanced after exposure to sub-inhibitory concentrations of florfenicol. These results suggest that cattle and swine can be treated with florfenicol for respiratory illness without exacerbating Salmonella Typhimurium DT104 virulence in carrier animals.

Detection of Salmonella Enteritidis in asymptomatic carrier animals: comparison of quantitative real-time PCR and bacteriological culture methods

Quantification of Salmonella in asymptomatic carrier animals can be used to assess microbial risk and monitor the level of contamination in domestic animals used for food production. We examined the sensitivity, specificity and accuracy of real-time qPCR, without pre-enrichment or selective enrichment stages, for the quantification of S. enterica serovar Enteritidis in resistant mice, as a model of asymptomatic carrier animal. The results were compared with those obtained by traditional bacteriological culture methods, the gold standard test. Two hundred and forty-three samples, including spleen, liver, mesenteric lymph nodes, portions of intestine, intestinal content of the ileocecal portion, and feces, were collected from a group of 27 C57BL/6 mice, that had been intragastrically inoculated with high doses of S. enterica serovar Enteritidis. The real-time qPCR assay presented a consistent linearity of the standard curve (r(2) = 0.999), with very low differences between melting temperatures, and low coefficients of variation in intra- (< 1%) and interassay (< 2%) comparisons. The primers were highly specific; there was no amplification with other Salmonella serovars or with DNA from uninfected tissues and feces from mice. The detection limit of the technique was defined as 32 copies of S. enterica serovar Enteritidis. A sensitivity of 90%, a specificity of 77% and an accuracy of 79% were obtained. The higher sensitivity of PCR was reflected in a kappa coefficient of 0.41, showing moderate agreement between tests. We conclude that real-time qPCR is a good alternative for diagnostic scanning in asymptomatic carrier animals, due to its high sensitivity and rapidity.

A Review of Bovine Anaplasmosis

Bovine anaplasmosis, caused by Anaplasma marginale, is an infectious but non-contagious disease. It is spread through tick bites or by the mechanical transfer of fresh blood from infected to susceptible cattle from biting flies or by blood-contaminated fomites including needles, ear tagging, dehorning and castration equipment. Transplacental transmission of A. marginale may contribute to the epidemiology of bovine anaplasmosis in some regions. Bovine anaplasmosis occurs in tropical and subtropical regions worldwide. Cattle of all ages are susceptible to infection with A. marginale, but the severity of disease increases with age. Once cattle of any age become infected with A. marginale, they remain persistently infected carriers for life. Diagnosis of bovine anaplasmosis can be made by demonstration of A. marginale on stained blood smears from clinically infected animals during the acute phase of the disease, but it is not reliable for detecting infection in pre-symptomatic or carrier animals. In these instances, the infection is generally diagnosed by serologic demonstration of antibodies with confirmation by molecular detection methods. The susceptibility of wild ruminants to infection by A. marginale and the role of wild ruminants in the epidemiology of bovine anaplasmosis are incompletely known owing to lack of published research, lack of validation of diagnostic tests for these species and cross-reaction of Anaplasma spp. antibodies in serologic tests. Control measures for bovine anaplasmosis vary with geographical location and include maintenance of Anaplasma-free herds, vector control, administration of antibiotics and vaccination.

HORSE SPECIES SYMPOSIUM: Contagious equine metritis: An insidious threat to the horse breeding industry in the United States1

Contagious equine metritis (CEM) has given rise to international concern since it was first recognized as a novel venereal disease of equids in 1977 and the etiologic agent was identified as a previously undescribed bacterium, Taylorella equigenitalis. Horse industry concerns over CEM centered on the ease with which this bacterium could be disseminated, the significance of T. equigenitalis as a cause of short-term infertility in the mare, and the existence of the carrier state in the stallion and the mare. The first known outbreak of CEM in the United States was in Kentucky in 1978. The economic impact on the Thoroughbred industry in the state was substantial. Before 2008, additional small-scale outbreaks occurred in Missouri in 1979, Kentucky in 1982, and Wisconsin in 2006, nearly all attributed to the importation of carrier animals. On each occasion, appropriate measures were taken to eliminate the infection, resulting in the United States regaining its CEM-free status. With the exception of the 1978 occurrence in Kentucky, none of the subsequent outbreaks significantly affected the horse industry. That changed dramatically in 2008, however, after the discovery of a Quarter horse stallion in Kentucky that cultured positive. Subsequent investigations turned up 23 carrier stallions and 5 carrier mares belonging to 11 breeds and located in 8 states. Shipment of infective semen and indirect venereal contact in stallion collection centers through the use of contaminated fomites were major factors in the spread of T. equigenitalis. Trace-back investigations of some 1,005 exposed and carrier stallions and mares in 48 states have failed to identify the origin of this latest CEM event. Neither clinical evidence of CEM nor decreased pregnancy rates were reportedly a feature in infected or exposed mares. In light of these findings, there was some question of whether or not the considerable expense incurred in investigating the latest CEM occurrence was warranted. Regaining CEM-free status for the United States will present considerable challenges.

Animal leptospirosis in small tropical areas

Leptospirosis is the most widespread zoonosis in the world. Humans become infected through contact with the urine of carrier animals, directly or via contaminated environments. This review reports available data on animal leptospirosis in ten tropical islands: Barbados, Martinique, Guadeloupe, Grenada, Trinidad, New Caledonia, Hawaii, French Polynesia, La Réunion and Mayotte. Leptospirosis is endemic in these insular wild and domestic fauna. Each island presents a specific panel of circulating serovars, closely linked with animal and environmental biodiversity, making it epidemiologically different from the mainland. Rats, mongooses and mice are proven major renal carriers of leptospires in these areas but dogs also constitute a significant potential reservoir. In some islands seroprevalence of leptospirosis in animals evolves with time, inducing changes in the epidemiology of the human disease. Consequently more investigations on animal leptospirosis in these ecosystems and use of molecular tools are essential for prevention and control of the human disease.

Evaluation of cytochrome b as a sensitive target for PCR based detection of T. annulata carrier animals

Bovine tropical theileriosis, caused by the tick-borne protozoan Theileria annulata, imposes a serious constraint upon breed improvement programmes and livestock production in tropical and sub-tropical regions of the world. Animals that recover from primary infection serve as carriers and play a critical role in the epidemiology of the disease, acting as reservoirs of infection. However, conclusive identification of carrier animals can be problematic. This study describes assessment of candidate target genes for PCR assay-based detection of T. annulata infected carrier animals. Following in silico screening and rejection of three major multi-copy gene families, an assay based on PCR amplification of a 312bp segment of the T. annulata gene for cytochrome b (Cytob1 assay) was established. Sensitivity was evaluated using serial dilutions of blood obtained from experimentally infected calves, while specificity was confirmed by testing DNA representing twelve different T. annulata stocks and other Theileria and Babesia species. Direct comparison with other target genes and published data indicated that Cytob1 PCR-based assays provide the greatest level of sensitivity, combined with a high level of specificity and the ability to detect different T. annulata genotypes. It can be concluded that the cytochrome b gene is the optimal target for PCR amplification and its incorporation in a Reverse Line Blot Assay offers the most sensitive method yet devised to detect the parasite in carrier animals. The use of this assay will increase the accuracy of epidemiological studies aimed at improving disease control in endemically unstable regions.

Asymptomatic Renal Colonization of Humans in the Peruvian Amazon by Leptospira

BackgroundRenal carriage and shedding of leptospires is characteristic of carrier or maintenance animal hosts. Sporadic reports indicate that after infection, humans may excrete leptospires for extended periods. We hypothesized that, like mammalian reservoir hosts, humans develop asymptomatic leptospiruria in settings of high disease transmission such as the Peruvian Amazon.Methodology/Principal FindingsUsing a cross-sectional study design, we used a combination of epidemiological data, serology and molecular detection of the leptospiral 16S rRNA gene to identify asymptomatic urinary shedders of Leptospira. Approximately one-third of the 314 asymptomatic participants had circulating anti-leptospiral antibodies. Among enrolled participants, 189/314 (59%) had evidence of recent infection (microscopic agglutination test (MAT0 ≥1∶800 or ELISA IgM-positive or both). The proportion of MAT-positive and high MAT-titer (≥1∶800) persons was higher in men than women (p = 0.006). Among these people, 13/314 (4.1%) had Leptospira DNA-positive urine samples. Of these, the 16S rRNA gene from 10 samples was able to be sequenced. The urine-derived species clustered within both pathogenic (n = 6) and intermediate clades of Leptospira (n = 4). All of the thirteen participants with leptospiral DNA in urine were women. The median age of the DNA-positive group was older compared to the negative group (p≤0.05). A group of asymptomatic participants (“long-term asymptomatic individuals,” 102/341 (32.5%) of enrolled individuals) without serological evidence of recent infection was identified; within this group, 6/102 (5.9%) excreted pathogenic and intermediate-pathogenic Leptospira (75–229 bacteria/mL of urine).Conclusions/SignificanceAsymptomatic renal colonization of leptospires in a region of high disease transmission is common, including among people without serological or clinical evidence of recent infection. Both pathogenic and intermediate Leptospira can persist as renal colonization in humans. The pathogenic significance of this finding remains to be explored but is of fundamental biological significance.

Prevalencia de Salmonelosis en avícolas tecnificadas de postura del Departamento del Huila

La infección de origen alimentario por Salmonella es de las causas más importantes de intoxicación alimentaria en humanos y animales. Los reservorios son animales portadores asintomáticos y las fuentes de infección son alimentos o productos de estos. El aumento de la incidencia de Salmonella es de gran impacto tanto en salud pública y salud animal principalmente por la diseminación de microorganismos en la industria avícola. En el mundo los indicadores de prevalencia son del 10% al 17% en humanos y del 25% al 55% en animales, incluyendo aves de engorde. El objetivo de esta investigación es identificar la prevalencia de salmonelosis, en avícolas de postura en la zona rural de los municipios de Neiva, Rivera y Palermo. Para tal fin se incluyeron aves de 19 a 68 semanas de edad para llevar a cabo este estudio de corte transversal. Se practicó la caracterización de cada granja, teniendo en cuenta diferentes aspectos: tamaño, línea genética, edad y peso de las aves; fueron evaluadas las condiciones de bioseguridad, y posteriormente recolectada la información con lista de chequeo. Para lograr definir la prevalencia de salmonelosis se realizó por parte de los médicos veterinarios zootecnistas la toma de frotis cloacal a 372 aves; se tomaron dos muestras: una para estudio y otra para control por ave; luego fueron llevadas al laboratorio del Instituto Colombiano Agropecuario, para su análisis; finalmente no fue posible la detección de la bacteria Salmonella en las muestras estudiadas provenientes de las cuatro granjas de estudio.

Molecular characterization of SAT 2 foot-and-mouth disease virus from post-outbreak slaughtered animals: implications for disease control in Uganda

In Uganda, limiting the extent of foot-and-mouth disease (FMD) spread during outbreaks involves short-term measures such as ring vaccination and restrictions of the movement of livestock and their products to and from the affected areas. In this study, the presence of FMD virus RNA was investigated in cattle samples 3 months after FMD quarantine measures had been lifted following an outbreak in 2004. Oropharyngeal tissue samples were obtained from 12 cattle slaughtered in a small town abattoir in Kiboga. FMD virus RNA was detected by diagnostic RT-PCR in nine of the 12 tissue samples. Part of the coding region for the capsid protein VP1 was amplified and sequenced. All samples were identified as belonging to the SAT 2 serotype. The implications for FMD control of both virus introduction into Uganda and the presence of carrier animals following outbreaks are discussed.

Differentiation of human adipose-derived stem cells induced by recombinantly expressed fibroblast growth factor 10 in vitro and in vivo

The adipogenesis effect of fibroblast growth factor 10 (FGF10) has been demonstrated in many studies. The aim of this study is to render a novel method which can continuously induce hypodermal adipose-derived stem cell (ADSC) differentiation and maturation in vivo and in vitro using FGF10. We constructed a recombinant pcDNA3.0-FGF10-MSC which can continuously express FGF10 by transfected FGF10 into a human mesenchymal stem cell (MSC) clone, and we cultured ADSCs from human subcutaneous resected adipose tissue. An in vitro and in vivo coculture system of pcDNA3.0-FGF10-MSC and ADSCs was then established. We observed the characteristics of ADSCs, monitored the adipogenesis-related transcription factor CAAT/enhancer binding protein-β, peroxisome proliferator-activated receptor-γ, and measured the adipose tissue layer of carrier animals. The results showed that FGF10 secreted from pcDNA3.0-FGF10-MSC could induce ADSC differentiation into mature adipocytes consistently. The study demonstrated that FGF10 can promote the adipogenesis effect in situ, and the autotransplantation of a carrier continuously secreting FGF10 may be utilized for increasing local subcutaneous adipose tissue in cosmetology.

A nested PCR assay exhibits enhanced sensitivity for detection of Theileria parva infections in bovine blood samples from carrier animals

Theileria parva causes East Coast fever, an economically important disease of cattle in sub-Saharan Africa. We describe a nested polymerase chain reaction (nPCR) assay for the detection of T. parva DNA in cattle blood spotted onto filter paper using primers derived from the T. parva-specific 104-kDa antigen (p104) gene. The sensitivity of this assay was compared to a previously described p104-based PCR and also the reverse line blot (RLB) technique, using serial dilutions of blood from a calf with known T. parva piroplasm parasitaemia. The relative sensitivities of the three assays were 0.4, 1.4 and 4 parasites/microl corresponding to blood parasitaemias of 9.2 x 10(-6)%, 2.8 x 10(-5)% and 8.3 x 10(-5)%, respectively. The three assays were applied to samples from two calves infected with the T. parva Muguga stock. Parasite DNA was consistently detectable by the two p104 PCR assays until 48 and 82 days post-infection, respectively, and thereafter sporadically. RLB detected parasite DNA in the two infected calves until days 43 and 45. Field samples from 151 Kenyan cattle exhibited 37.7% positivity for T. parva by regular p104 PCR and 42.3% positivity using p104 nPCR. Among 169 cattle blood samples from Southern Sudan, 36% were positive for T. parva using nPCR. The nPCR assay represents a highly sensitive tool for detection and monitoring of asymptomatic carrier state infections of T. parva in the blood of cattle.

Salmonella enterica: isolamento e soroprevalência em suínos abatidos no Rio Grande do Sul

Estudou-se a prevalência de Salmonella spp. em suínos ao abate e compararam-se os resultados obtidos no isolamento e na sorologia de modo a determinar a fase mais importante de infecção (granja ou transporte/espera pré-abate) em relação ao número de suínos portadores encontrados ao abate. Foram avaliados 40 rebanhos de três diferentes empresas do sul do Brasil, com colheitas de sangue e linfonodos mesentéricos de 20 animais por lote. O soro foi submetido ao ELISA produzido com antígeno somático de Salmonella Typhimurium. A frequência de isolamento variou de 62,5% a 85,0%, enquanto a soroprevalência de 73,8% a 83,2% nos três sistemas amostrados. Os sorovares mais prevalentes foram Agona, Typhimurium e Panama. A infecção dos animais ocorreu nas granjas produtoras de suínos nestes sistemas de produção, pois os lotes apresentaram elevada soroprevalência ao abate. Concluiu-se que o controle da infecção por Salmonella spp. no sul do Brasil deve iniciar-se pela implementação de medidas de profilaxia nas granjas de produção de suínos.

Monitoring of Lawsonia intracellularis in breeding herd gilts

In modern pig production, proliferative enteropathy is a common cause of diarrhoea and poor growth in young animals. This study aimed to determine the possible spread of Lawsonia intracellularis through the sale of replacement gilts and the possibility to protect the herds by adequate biosecurity measures. This was achieved by repeated sampling of 50 gilts in an infected multiplying herd, from the last day in the farrowing pen and until sale. Further, 60 gilts sold from this herd were tested during their stay in quarantine in a recipient herd. To confirm freedom from infection, 100 growing pigs in the recipient herd were also tested. Individual faecal (n=748) and blood (n=728) samples were analysed by PCR and ELISA, respectively. Transmission of L. intracellularis from the sows to their offspring was not demonstrated. However, the possible transmission between herds by replacement gilts was demonstrated. Peak shedding occurred at 12 and 15 weeks of age, and single animals were also PCR-positive at 24–36 weeks of age in the multiplying herd and in the quarantine in the recipient herd. Further, the possible occurrence of chronically infected carrier animals was suggested. Although L. intracellularis is widely spread, it appears possible to avoid the transmission between herds by employing adequate biosecurity measures. Thus, it would be advisable to establish herd profiles in breeding herds to avoid the selling of infected animals as well as to establish the health status of the recipient herd. Further, the health status of the recipient herds should be known.

Field validation of a competitive ELISA for detection of Theileria annulata infection

In previous studies, Theileria annulata surface protein (TaSP) was identified as an immunodominant antigen and successfully used to develop a recombinant-protein-based indirect enzyme-linked immunosorbent assay (ELISA) for the detection of circulating antibodies in serum of T. annulata-infected animals. To increase the specificity, a competitive ELISA (cELISA) was developed using recombinant TaSP antigen and a monoclonal antibody (1C7) specifically binding to TaSP. Since the cELISA accurately differentiated T. annulata-infected from uninfected animals, a study was performed to analyse the suitability of the cELISA in the field. For this, 230 sera with unknown status from different governorates in the north of Iraq were analysed using both the indirect and competitive ELISA and were compared. There was a significant (p < 0.5 x 10(-19)) correlation (r = 0.556) between the tests, whereby the cELISA detected more sera as negative (44/230) compared to the indirect ELISA (21/270). Accordingly, less sera were determined to be positive in the competitive (186/230) than in the indirect ELISA (209/230). Sensitivity and specificity of the cELISA taking the indirect ELISA as a reference were 84.2% and 52.4%, respectively. Accordingly, the calculated prevalence of T. annulata infection was 90.9%, and the positive predictive value was determined to be 94.6%. Taken together, the cELISA proved its suitability for field application and was found qualified for use in serological surveys to monitor the prevalence of T. annulata infection and to identify carrier animals.

Eradication of feline dermatophytosis in a shelter: a field study

Enzootic dermatophytosis in a shelter with approximately 140 cats was treated according to a protocol combining identification, isolation and treatment of subclinical carrier and affected animals in accordance with a three-area system: healthy animals (no lesions and negative cultures), subclinical carrier animals (no lesions but with positive cultures) and clinically affected animals (lesions and positive cultures). The cats were examined and inspected under a Wood's lamp and had samples taken for fungal culture every 2 weeks. Thirty-three per cent of the cats had a positive fungal culture at the start of the study. Clinically affected animals and carriers were treated with a 0.2% enilconazole lotion (Imaverol) twice a week and given itraconazole (Itrafungol) 5 mg/kg SID orally every other week. The environment was treated once a day with a 1% bleach solution and once a week with a 0.6% enilconazole (Clinafarm) solution. Treated animals were considered cured after two consecutive negative fungal cultures. All cats were cured within 56 days. Prophylactic measures against dermatophytosis were implemented for new arrivals consisting of individual quarantine and the systematic taking of fungal cultures. No relapses were observed based on the fungal cultures taken from the animals and the environment over the first 10 months.

Influence of Exogenous Melatonin on Horizontal Transfer of Escherichia coli O157:H7 in Experimentally Infected Sheep

The objective of the current research was to determine if exogenous melatonin would exert a "protective" effect on the gastrointestinal tract of sheep and prevent or reduce the horizontal transfer of Escherichia coli O157:H7 from experimentally infected to noninfected or "naïve" sheep. Sixteen cross-bred ewes were housed indoors and adapted to a high concentrate ration. Ewes were randomly assigned to one of four rooms and treatment (three ewes/room, six ewes/treatment) and received either control (gelatin capsule only) or melatonin (5.0 mg/kg body weight [BW]/d). Four additional ewes served as "carrier" sheep (one/room) and were experimentally infected via oral gavage with E. coli O157:H7. Three days post-challenge, carrier ewes were housed with naïve sheep and remained with them for the remainder of the experimental period. Treatments were administered to the naïve sheep 1 day prior to introduction of the carrier sheep and on each of the remaining 7 days of the experimental period. Fecal samples were collected via rectal palpation from the carrier sheep daily throughout experiment and from the naïve sheep daily for 5 days, starting 2 days following introduction of the carriers. On day 8 of the experiment, all ewes were euthanized and tissues from the rumen, ileum, cecum, colon, and rectum as well as their respective lumen contents collected. The carrier sheep quickly infected the naïve ewes, which had similar fecal concentrations as the carrier animals throughout the 5-day sampling period. Melatonin treatment had no effect (p > 0.10) on daily fecal shedding, luminal content concentrations, or in the percentage of gastrointestinal tract tissue positive for the inoculated strain of E. coli O157:H7.

Protective vaccination in the horse against Streptococcus equi with recombinant antigens

AbstractStreptococcus equi subspecies equi (S. equi) is a clonal, equine host-adapted pathogen of global importance that causes a highly contagious suppurative lymphodendopathy of the head and neck, more commonly known as Strangles. The disease is highly prevalent, can be severe and spread easily by visibly infected animals or by carrier animals that show no clinical signs of disease. Antibiotic treatment is usually ineffective. However, the majority of horses develop immunity to re-infection, suggesting that vaccination should be a feasible way to prevent the infection. Live attenuated vaccine strains of S. equi are available but adverse reactions have been reported and they suffer from a short duration of immunity. Thus, a safe and effective vaccine against S. equi is highly desirable. In this report, Welsh mountain ponies vaccinated with a combination of seven recombinant S. equi proteins, were significantly protected from experimental infection by S. equi, resembling the spontaneous disease. The protective antigens consisted of five surface localized proteins and two IgG endopeptidases. The results from a second vaccination trial indicate that the endopeptidases were important for good protection. The similarity of S. equi to other pyogenic streptococci suggests that our findings have broader implications for the prevention of streptococcal infections.

The effect of vaccination on undetected persistence of foot-and-mouth disease virus in cattle herds and sheep flocks

The importance of carrier animals (those in whom virus persists after recovery from disease or acute infection) and their potential role in the spread of disease remain open questions within foot-and-mouth disease epidemiology. Using simple probabilistic models we attempt to quantify the effect of emergency vaccination--and especially the time of application--on the likely number of such animals, using data from challenge experiments on both cattle and sheep to determine the probability of persistence in diseased and subclinically infected animals. We show that the number of persistently infected animals in a group is predominantly determined by the number of animals initially infected on premises--the high variability of which ultimately limits the accuracy of any predictions of carrier numbers based upon transmission models. Furthermore, results suggest that, within a cattle herd, carrier numbers may be increased if challenge occurs shortly after vaccination. We show that the quality of inspection is the principal factor influencing whether or not carrier herds occur and that, by reducing clinical signs, the application of vaccination in regularly checked stock also results in an increase in undetected persistently infected animals. Where clinical detection would be poor regardless of the use of vaccination (i.e. particularly in sheep), vaccination will result in a reduction in the probability of a group containing undetected carriers: otherwise there is a benefit only if vaccination is applied sufficiently far in advance of any challenge. The implications of the results for serosurveillance are discussed, including the requisite test sensitivity and practices for successful implementation.

Identification of Leptospira spp. carriers among seroreactive goats and sheep by polymerase chain reaction

Few studies were conducted on the diagnosis and control of small ruminants’ leptospirosis. Thirteen goat herds and seven sheep flocks located in the state of Rio de Janeiro, Brazil, were screened for leptospirosis. From the three herds and three flocks with greatest seroreactivity by MAT (Microscopic Agglutination Test), 19 and 40 seropositive goats and sheep, respectively, were selected, and urine samples were collected for bacteriology and PCR. For both species of animals, the most prevalent reactions were due to serogroups Sejroe and Shermani. Although leptospires were observed by darkfield microscopy in eight samples, pure isolates were obtained by bacteriological culture from only two samples. However, twelve urine samples (six goats and six sheep) were positive by PCR. Based on these findings, we consider that the combined use of MAT as a screening test followed by urine PCR for the direct detection of Leptospira spp. DNA was adequate for the identification of carrier animals among goats and sheep. These are valuable tools for the control of leptospirosis in small ruminants.