PCR-RLFP characterization of Leishmania spp. in domestic animals from the south-western border of Brazil.

The aim of this study was to characterize Leishmania spp. from canine and feline samples using Polymerase Chain Reaction (PCR)- Restriction Fragment Length Polymorphism (RFLP). It was conducted in the southern region of Brazil, located at border crossings to Argentina and Uruguay. Samples were collected from 116 dogs (Canis lupus familiaris) and 89 cats (Felis catus). The PCR was performed to screen for an LT1 fragment from kinetoplast DNA (kDNA) target gene, and positive samples were subjected to a second PCR for an internal transcribed spacers (ITS1) region from ribosomal DNA (rDNA) target. RFLP was performed using the Haemophilus aegyptius (HAE III) restriction endonuclease (Fermentas ®). Positive samples by PCR ITS1 were sequenced and deposited in NCBI GenBank, and a phylogenetic analysis was developed. We found that 12.9% (15/116) of the samples from dogs were positive. All the 89 cat samples were negative. Positive samples were tested against Leishmania reference strains presenting different patterns in PCR-RFLP, and these samples showed bands denoting similarity to the standard species of Leishmania infantum, proven through sequencing and phylogenetic analysis. The RFLP technique, alone, was shown to be feasible for practical application and confirmation of the involved Leishmania spp.

Risk and protective factors of Leishmaniasis in the rural area of the western border region of Rio Grande do Sul, Brazil.

BACKGROUND: The Leishmaniases are on the top of the global list of tropical neglected diseases. The number of infected dogs in South America is estimated in millions and correlated to disease cases in humans, especially in Brazil. Equines may get infected too and can play a role in the epidemiological chain. Thus, the aim of the present study was to evaluate risk and protective factors of leishmaniasis in rural areas of the western border region of Rio Grande do Sul state, Brazil by Leishmania spp. protozoa molecular detection and serological evaluation (ELISA) in equine and canine blood samples. This work included nine farms around the city of Uruguaiana. Epidemiologic information regarding farm characteristics and biologic material collection of canine (22) and equine (91), totalizing 113 samples was collected. The polymerase chain reaction (PCR) technique was used to detect Leishmania spp. in biological samples. Variables related to the farm were collected and evaluated through descriptive analysis followed by chi-square test and a logistic regression analysis. RESULTS: Nineteen positive samples (19/113 - 16,81%) were detected, being 18 equines and 1 canine, in six of the nine farms included in the study. No animal showed clinical signs of the disease. According to the variables analyzed, when compared each characteristic separately, the presence of abundant vegetation and poor hygiene demonstrated to be risk factors to Leishmania infection in rural areas. The logistic regression showed that excellent general hygiene, proximity to the weir and trimmed grass were protective factors (p=0.038, p=0.001 and p=0.014, respectively). Having excellent hygiene represents a 70% lower chance of getting infected, keeping the grass cut protects the animal by more than 90% and the proximity of the weir represents a protective factor of 96%. CONCLUSIONS: The presence of Leishmania infection in the western border region of Rio Grande do Sul was 16,81% and it was influenced by farm characteristics. The role of the excellent general hygiene as a protective factor is extremely relevant in the leishmaniases prevention.

Influence of morphometric parameters on the size of collateral ligaments of the distal interphalangeal joint of equines.

The physiological measurements of collateral ligaments of distal interphalangeal joint (CL-DIPJ) differ in the literature. The factors that influence these differences are not well described. The aims of this study are to compare CL-DIPJ sizes in equines with different withers height, as well as to correlate body weight and hoof size to the size of these ligaments. In total, 52 horses were used in the study. They were divided into two groups according to wither height: Group 1 (G1) - 21 animals with up to 147 cm - and group 2 (G2) - 21 animals with greater than 148 cm. CL-DIPJ was ultrasonographically measured in order to find the mean of dorso-palmar (DPD) and latero-medial (LMD) diameters and the cross-sectional area (CSA). Hoof width and length were measured, and the results were used to calculate the hoof surface area. Withers height and body weight were also measured. Groups were statistically compared by Student's t test and Pearson's correlation application to each group. Groups were different in body size, HS and CL-DIPJ size when p < .05, except for the LMD of the medial collateral ligament of left thoracic limb. Withers height and body weight did not show significant correlations to CL-DIPJ size in G1, but they had little influence on the size of CL-DIPJ on G2. No correlation between the hoof size and the CL-DIPJ was noted in any of the two groups. In conclusion, the CL-DIPJ were larger in taller horses and their sizes were correlated to their height and weight, but they were not correlated to hoof size.

Ceftiofur Side Effect in a Mare-Case Report.

Adverse drug reactions in horses are rare. The antimicrobials are in the list of the most common drugs associated with reaction in horses. The aim of this report is to describe the clinical presentation of an adverse drug reaction after the intramuscular administration of ceftiofur hydrochloride. A 5-year-old crioulo broodmare at one day postpartum presented signs of difficulty to walk, positive pulse in the four limbs, and heart rate of 80 beats per minute (bpm), with a history of fighting with another mare. The clinical suspicions were rhabdomyolysis and laminitis. Initially a dose of flunixin meglumine was administrated intravenous to prevent an endotoxemia. On the same day, the blood collection showed an elevated number of white blood cells, predominantly neutrophils and monocytes. An antimicrobial treatment with ceftiofur hydrochloride at a dose of 2.2 mg/kg intramuscular was applied. At the first day, the mare does not show signs of drug side effect. However, 24 hours later, in the second application, the patient presents incoordination, dizziness, and loss of equilibrium. At the same time, dexamethasone was administrated. The signs were reverted, and the mare was normal after the reaction. The treatment with ceftiofur was changed to enrofloxacin, and the animal had a complete recovery. These drug reactions are not common in the routine of equine clinical practitioners, but they cause a great preoccupation for the owner and the veterinarian.