Infection and re-infection of Leptospira spp. in stray dogs and cats from Bogota, Colombia.

Background and Aim: Leptospirosis is a re-emerging zoonosis that is under-reported in tropical countries, and canines can be a potential reservoir of the disease. The objective of this study was to diagnose Leptospira spp. that is actively infected and re-infected in stray dogs and cats from Bogota, D.C., Colombia. Materials and Methods: A sample of 200 animals, including dogs and cats from the animal protection programs of Bogota, Colombia, were used in this study. Blood was collected from these animals for serum and DNA analysis. Conventional polymerase chain reaction (PCR) was performed using the 16s rRNA primer set, and higher-quality amplification products were sequenced by Sanger. For serodiagnosis, a group of PCR-positive samples was tested using the microagglutination test (MAT). Results: The overall PCR positivity of stray dogs and cats was 56%, 52.9%, and 65.3% in dogs and cats, respectively. The MAT seropositivity was 77.3%, and only dogs showed titers higher than 1:400. Canicola, Icterohaemorrhagiae, Pomona, Hardjo Prajitno, and Canicola and Hardjo prajitno were the serogroups associated with dogs and cats, respectively. Phylogenetic analysis revealed that the strains belonging to Leptospira interrogans serovars related to isolated samples of American, European, and Asian bats (Myotis myotis), dogs, and bovines of American origin. Conclusion: These results showed that stray dogs and cats were previously exposed to different serovars of Leptospira spp. and re-infected with other serovars that actively participated in the transmission cycle. These findings highlight the importance of actively diagnosing infectious animals to design effective intervention strategies.

Seroprevalence and risk factors for Leptospira spp. in small ruminants of semi-arid zone in northeastern Colombia.

Leptospirosis in small ruminants is an economically important disease that causes important losses by pathologies of the reproductive tract. The present study aimed to estimate the seroprevalence and risk factors associated with Leptospira spp. in small ruminants under traditional husbandry systems. A cross-sectional epidemiological study was carried out in a sample of 1039 animals which included 793 sheep and 245 goats from 49 farms of 10 municipalities of northeastern areas of Colombia. The MAT test using a panel of 13 laboratory serovars was performed to diagnose the Leptospira spp. antibodies in the serum samples of animals. A structured questionnaire was filled in each farm to obtain information on putative risk factors, and its association with Leptospira spp. infection was detected by univariate and multivariate logistic regression. The general seroprevalence showed a rate of 13.9%, the 79.6% of farms showed at least one positive animal, the 14.2% and 0.4% of positive animals showed titers ≥ 1:100 and 1:200, respectively. The seroprevalence in sheep and goats showed a rate of 13.1% and 16.2%, respectively. The most common serovars in positive animals were Autumnalis (3.6%), Tarassovi (3.4%), Bratislava (2.9%), Mini (2.9%), Bataviae (1.4%), Canicola (0.3%), and Grippotyphosa (0.3%). The univariate and multivariate analysis showed as risk factors the absence of housing system (OR = 3.412 CI: 1.347-8.152), the absence of quarantine measures to new animals (OR = 1.371 CI: 0.943-1.978), and the protective factor identified was the use of Toggenburg breed (OR = 0.571 CI: 0.249-1.074). This exploratory study showed the Leptospira spp. positive small ruminants located in the northeastern areas of Colombia with titer associated with chronic and subclinical infection.

Cattle seroprevalence and risk factors associated with bovine viral diarrhea in the northeastern of Colombia.

Bovine viral diarrhea (BVD) is one of the most common and economically significant viral infections in cattle. Different risk factors have been associated with viral exposure and transmission, generating endemic regions with different biotype circulations. To find the BVD status in the northeastern region of Colombia, this study aimed to determine the seroprevalence and risk factors associated with the disease in non-vaccinated farms. For this purpose, a two-time point sampling strategy was developed, obtaining serum from 1157 animals housed in 25 farms distributed in 46 townships, thus for antibody detection against non-structural protein 3 (NS3, p80) of bovine viral diarrhea virus (BVDV) using an indirect ELISA test. For the first time in Colombia, the presence of serological persistent cattle was evaluated in 1047 animals 3 weeks later of first sampling. The information on a standardized questionnaire with closed and dichotomic answers was used to calculate the putative risk factors. The association analysis by univariate and multivariate logistic regression reported odds ratios (OR) with a 95% of confidence interval (C.I). The overall seroprevalence was 65% (95% CI 62.5-72.9%), with variations according to age ranges. The prevalence of persistent serological positive corresponded to 37.7% (95% CI 34.2-41.5). The risk factors found by multivariate analyses were the lease of pastures (OR = 2.071 CI 1.485-3.690), the use of the same needle (OR = 2.249 CI 1.354-3.736), the molasses supplementation (OR = 2.742 CI 1.156-5.807), and the native Creole breed (OR = 1.895 CI 1.416-2.804). The results of this study confirmed the endemism and higher common exposure to BVDV, as well as the presence of serological persistent cattle in Valledupar, Colombia.

Bovine immunisation with a recombinant peptide derived from synthetic SBm7462® (Bm86 epitope construct) immunogen for Rhipicephalus microplus control.

RHIPICEPHALUS MICROPLUS: is the most important ectoparasite of livestock in tropical and subtropical areas around the world. Research focused on developing an efficient vaccine for cattle tick control is a high priority. The aim of this study was to evaluate the rSBm7462® peptide (Bm86-B and T cell epitopes) regarding its properties of immunogenicity, protective effect in cattle and efficacy against R. microplus. This peptide was produced by a fermentative process in the yeast culture system of Komagataella (Pichia) pastoris strain Km 71. The vaccination assay was conducted in a tick-free area using non-splenectomised Holstein Friesian calves, separated into immunisation and control groups. These animals individually received the recombinant peptide and the inoculum without peptide using saponin as an adjuvant at three time points. The calves were challenged 21 days after the last immunisation using 4500 larvae per animal. An indirect ELISA was used to identify the IgG kinetics of serum samples from the calves studied. The qPCR was performed to determine the cytokine gene expression from the total RNA of the cultured PBMCs. Histomorphometry of the germinal centres (GCs) was performed measuring slides with haematoxylin-eosin staining of surgically removed lymph nodes from immunised calves. The antibody response showed a significant induction of high-affinity IgGs in calves immunised with the recombinant peptide in comparison to the controls. The kinetics of antibodies in immunised calves showed a significant increase during the experiment. This increase in high-affinity IgGs correlated with a gradual increase of the GC diameter following each peptide vaccination. Cytokine expression profiles demonstrating an adaptive immune response in calves immunised with rSBm7462® confirmed the T-dependent response. Vaccine efficacy was calculated at 72.4 % following the analysis and fecundity of collected adult female ticks, compared between control and vaccinated groups. These findings demonstrate that this new recombinant peptide is an option for control of R. microplus infestations.

Toxoplasma gondii in small ruminants in northeastern areas of Colombia: Seroprevalence and risk factors.

Sheep and goats are susceptible to infections with Toxoplasma gondii and could play an important role in the transmission of the zoonotic parasite to human. We conducted a cross sectional study to estimate the seroprevalence and to assess the risk factors for T. gondii seropositivity in small ruminants under traditional husbandry systems. This study was carried out from November 2015 to April 2016 in randomly selected small ruminants (n = 1038) from 48 farms located in Colombia, in the departments of northern Cesar in the north and La Guajira in the south. An indirect ELISA was used to detect IgG antibodies to T. gondii in the animals. A standardized questionnaire was used to obtain information on putative risk factors. We conducted the association analyses by using univariable and multivariate logistic regression and report odds ratios (OR) with 95% confidence interval (C.I). The overall seroprevalence in small ruminants was 23.5% (C.I: 21-26.2%). Sheep showed a higher seroprevalence (25.1% C.I: 22.4-28.6%) than goats (18.4% C.I: 22.4-28.6%). The association analysis recognized as risk factors for T. gondii seropositivity farming pigs in addition to small ruminants (OR = 1.96 C.I: 1.414-2.743), the inexistence of manure heap (OR = 2.254 C.I: 1.480-3.433) and drinking water from locally aqueducts (OR = 1.489 C.I: 1.006-2.204). The results of the study confirmed that exposure to T. gondii is common in sheep and goats in dry Caribbean regions of Colombia.