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ABSTRACT: Acute abdomen cases are among the most prevalent and challenging illnesses in equine medicine. The diagnosis includes anamnesis and detailed clinical evaluation of the patient, allowing the gathering of several important information and, additionally, the use of complementary exams helps to define the diagnosis and prognosis of each case. This research applied the multiple correspondence analysis (MCA) in data referring to clinical examination information and outcome of horses with colic referred to a hospital in Rio Grande do Sul, Brazil. Fifty eight horses were included in the analysis and, it was possible to identify significant differences between the groups, survivors and non-survivors. Related to clinical signs, non-survivor cases are distinguished by severe pain, lack of motility, mucosal alteration, capillary refill time greater than 2sec, heart rate more than 48bpm and presence of nasogastric reflux. As complementary exams, hematocrits and blood lactate demonstrate significative difference among the groups. The cardiovascular parameters, including heart rate, mucosal color and capillary refill time, were the variables with the greatest contribution to discriminate animals that non-survive from those that were discharged. Thus, clinical evaluation should not be underestimated and complementary exams should be used in conjunction to seek a better prognosis. The MCA allowed to visualize the data and predict the outcome of the patients.

ABSTRACT: Feline leishmaniosis is infrequent worldwide, and cats have been suggested as secondary reservoirs for the parasite. However, specific diagnostic techniques for feline samples are scarce. In this study, we standardized an in-house indirect enzyme-linked immunosorbent assay (ELISA) using crude Leishmaniainfantum antigen to detect antibodies in feline samples from an endemic canine visceral leishmaniosis (CVL) area in the western border of Brazil. The results were compared with those of an indirect immunofluorescence assay (IFA). We tested semi-domiciled felines residing in Uruguaiana and Barra do Quaraí, Rio Grande do Sul. Among the 41 samples, 25 (61%) were positive using ELISA and 24 (58%) were positive using IFA (1:40). Our findings demonstrated a high seropositivity of feline samples from the endemic CVL area in the western border of Brazil, and we proposed the use of an in-house ELISA with crude antigen for population screening. This is the first serological survey on felines in a region where CVL is well established.

Abstract 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.

ABSTRACT: Leishmaniosis is a great public health problem affecting both humans and animals. The disease is caused by the protozoan Leishmania spp., which has a complex cycle involving a phlebotomine vector. The ELISA test (Enzyme-Linked Immunosorbent Assay) along with a chromatographic immunoassay was defined by the Brazil Health Ministry as the confirmatory screening protocol in 2011. Uruguaiana city is 630 km away from Porto Alegre, which makes it difficult to send samples and diagnose leishmaniasis, as well as receive quick results. In view of this, the present study evaluated an in-house indirect ELISA method compared to indirect immunofluorescence assay (IFA) and dual-path platform chromatographic immunoassay (DPP-BioManguinhos®) for the detection of an immune response to Leishmania spp. in canine species. The serological evaluation included 48 canines from the western border of Brazil (Uruguaiana and Barra do Quaraí city). Among the 48 canine samples tested, 18 were positive when using the ELISA technique, 19 were positive with IFA, and 17 were positive with rapid test DPP®. The ELISA technique showed a sensitivity/specificity of 83.3%/86.7% when compared to IFA and 100%/96.8% compared to DPP®. The present study showed a prevalence of 37.5%, demonstrating that the infection circulates in the studied population. It can be concluded that the ELISA technique was valuable for use in field conditions when performing screening tests in endemic areas.

Abstract 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.

Abstract Background Leishmaniosis, zoonosis that produces significant public health impacts, is caused by Leishmania infantum. Canines are the main domestic reservoir and, besides humans, other species of mammals could be infected when living in endemic areas. In this study, we detected equine Leishmania infantum infections in a canine visceral leishmaniosis transmission area and evaluated the clinical, haematological, biochemical and oxidative stress disorders. This study was conducted in Uruguaiana, Rio Grande do Sul, south of Brazil. Peripheral blood samples were collected from 124 animals (98 horses and 26 dogs) of both genders and several breeds after they underwent general and dermatologic examinations. Results Twenty five Leishmania infantum infected animals (20.16%), 14 horses and 11 dogs were detected by PCR (Polymerase Chain Reaction) amplification of kinetoplast DNA regions with 96% homology to Leishmania infantum (GenBank Accession No. L 19877.1). The clinical and haematological alterations of infected equines were skin lesions, nodules, lymphadenopathy, decreased levels in red blood cells and haematocrit (p 0.05) in both species. Conclusions To our knowledge, this has been the first leishmaniosis equine survey performed in south of Brazil, caused by Leishmania infantum that were able to initially identify haematological and biochemical changes in the species, even in asymptomatic animals. We present evidence supporting those findings of haematological and biochemical changes could be related to infection. Surprisingly, the clinical manifestations of equine infection were similar to those found in canine visceral leishmaniosis. The equine population could be play an important role in the cycle of leishmaniosis in south Brazil and consequently indicates a great risk of public health. This evaluation of infected animals is important to establish the clinical and laboratory parameters involved in the disease progression.

Visceral leishmaniasis (VL) is a zoonosis caused by the protozoan of the genus Leishmania. The disease is transmitted by the bite of a sand fly vector. Although the main reservoirs are dogs, other hosts can be infected and may play this role. Rio Grande do Sul western region, located on the triple border of Brazil-Uruguay-Argentina, represents a VL transmission area. The goal of the present study was to identify Leishmania spp. infection in animals from rural areas of Uruguaiana, Rio Grande do Sul. Nine farms in the Uruguaiana municipality, Rio Grande do Sul state, were included. Peripheral blood samples were collected from 113 animals (canine [n=22], equine [n=91]) for detection of Leishmania spp. DNA was isolated and polymerase chain reaction was performed. Eight (7%) animals with Leishmania spp. infection were detected on two farms in the same geographical area, seven of which were horses and one was canine, all of which were asymptomatic. To investigate the species of Leishmania, one of the positive equine samples was subjected to direct sequencing, which confirmed the presence of L. infantum genetic material. Results of this study confirm the presence of L. infantum-infected animals in rural areas of Uruguaiana, and provide evidence supporting further investigation of risk factors for dissemination in such areas.

The present study aimed to evaluate factors associated with canine visceral leishmaniasis (CVL) in areas with higher seroprevalence of it in Uruguaiana city, Rio Grande do Sul state, Brazil, during the year of 2012. A semi-structured questionnaire with socioeconomic and environmental questions was applied in Leishmania endemic regions of the city. The survey data were analyzed by multivariate statistics and the associations between them were presented as odds ratio within a 95% confidence limit. The relationships between the studied variables showed no statistically significant difference between the dwellings with positive and negative dogs. However, when analyzing the odds ratio, the presence of green areas larger than 10m² turned the house into an area of potential risk for CVL (OR= 2.53). There was no difference among the socioeconomic variables education and income, though the groups with lower education and income showed a higher seroprevalence of CVL. The city of Uruguaiana is already located within the CVL broadcasting area and is taken as an endemic region of the disease, with seropositive dogs in all neighborhoods. Thus, the determinants for an increased Leishmania sp. infection could not be indicated, even so, the maintenance of green areas around the households represented a risk factor for being a potential vector shelter.

A domesticação dos animais, ocorrida há muitos anos, intensificou a relação de dependência homem-animal. Esta proximidade resultou em um crescente número de animais de companhia e conseqüente aumento do número de animais errantes nos centros urbanos, minando os preceitos de bem estar animal e ocasionado problemas de saúde pública. O município de Uruguaiana enfrenta grave problema de saúde pública devido ao elevado número de animais abandonados nas ruas. O presente trabalho objetivou avaliar a conscientização de adolescentes entre 13 a 16 anos do ensino fundamental sobre a guarda responsável de animais antes e após as ações do Projeto “Melhor Amigo”. Para tal foram utilizados um vídeo educativo e palestra, ambos abordando conceitos básicos de bem estar animal, controle de natalidade, prevenção de zoonoses e adoção responsável. Antes e após as apresentações foram aplicados questionários objetivos aos alunos para se avaliar a eficácia das apresentações. Observou-se que 87,3% dos adolescentes possuem animais de estimação, justificando a necessidade do projeto. Foram obtidos resultados estatisticamente significativos na melhora da conscientização dos alunos com relação aos quesitos: manutenção do animal nos limites da moradia, dieta mais adequada ao animal e formas de prevenção da transmissão da toxoplasmose. Os adolescentes demonstraram ter boa noção dos compromissos básicos de um proprietário consciente para com seu animal: alimentação, vacinação, castração e dedicação, mesmo antes dos trabalhos do projeto. E não houve alteração estatística no entendimento da leishmaniose, sendo necessária adequação metodológica na abordagem do assunto em trabalhos futuros.

Current analysis investigated the tickcide effects of the aqueous extract and chloroform fractions of Ruta graveolens L. (rue) on engorged females of Rhipicephalus microplus, as well as their genotoxic and mutagenic effects on human leukocytes. The best tickcide activity (non-dependent dose) and genotoxic / mutagenic effects (dependent-dose) were observed on exposure to chloroform fractions. Results suggest that extract fractions of R. graveolens L are efficient against R. microplus, although the fraction and the tested concentrations show genotoxic and mutagenic potential for human leukocytes.

A pitiose, doença granulomatosa de eqüinos causada pelo oomiceto Pythium insidiosum, tem como característica a evolução rápida seguida de morte dos animais. Estas mortes muitas vezes são causadas por diagnósticos errôneos ou demorados quando os doentes já não respondem ao tratamento. Este trabalho teve por objetivo a padronização do ensaio imunoenzimático indireto (ELISA) para diagnóstico sorológico de pitiose em eqüinos e coelhos, visando a diminuição de erros e de tempo necessário para o diagnóstico. Para o desenvolvimento e validação do teste foram utilizadas 72 amostras de soro de eqüinos saudáveis e 44 soros de eqüinos com pitiose confirmada. Os resultados da validação do ELISA para eqüinos foram: sensibilidade 97,72%, especificidade 90,27%, valor preditivo positivo 86%, valor preditivo negativo 98,4% e eficiência de 93,1%. Para coelhos, o teste foi padronizado com 48 amostras de soro de animais saudáveis e 24 amostras de coelhos imunizados com antígenos de P. insidiosum. Os resultados foram: sensibilidade 91,66%, especificidade 95,83%, valor preditivo positivo 91,66%, valor preditivo negativo 95,83% e eficiência de 94,44%. Os resultados deste trabalho demonstram que o ensaio imunoenzimático indireto é um método seguro e eficaz para o diagnóstico sorológico da pitiose.

Miscrosporum nanum is a dermatophyte found in swine that causes non-pruritic lesions with desquamation, alopecia, and circular characteristics. M. nanum infection in dogs is rare and poorly understood in terms of its epidemiological and clinical features, and its therapeutic response. The present report describes a case of dermatophytosis due to M. nanum in a Dogo Argentino breed of dog that was used for wild boar hunting. The dermatophytosis presented with hypotrichosis, erythema, and non-pruritic desquamation in the back of the neck and chest area. The dermatophytosis was responsive to systemic treatment with itraconazole and topical (miconazole 2%) for 60 days. Thus, we conclude that the practice of hunting wild boar should be considered as a possible source of infection of M. nanum in the reported dog. The M. nanum infection showed clinical features that were similar to the lesions observed in swine, except for the absence of the circular pattern, and showed a good clinical response to the therapy. Finally, M. nanum should be considered as an etiologic agent of dermatophytosis in dogs that in some manner have had direct contact with domestic or wild swine.

Leishmaniasis is a neglected zoonotic disease caused by a variety of pathogenic Leishmania species. In the New World, especially in Brazil, visceral leishmaniasis (VL) is caused by Leishmania infantum. The pathogen can infect several animal species including dogs, foxes, rodents, primates, felines, equines and humans. Dogs act as the primary domestic reservoirs. This study aimed to use polymerase chain reaction (PCR) for detecting Leishmania infection in horses living in a canine visceral leishmaniasis (CVL) endemic region. DNA samples from horse peripheral blood were used to perform PCR. Templates were amplified using primers for the kinetoplast DNA (kDNA) minicircles, which were able to detect different species of Leishmania. In addition, primers for internal transcribed spacer (ITS) of ribosomal DNA were used for detection of Trypanosomatidae sp. Amongst the 75 (39%) positive PCR samples from total 192 samples, 21 samples were positive for kDNA and 63 samples were positive for either ITS, ITS1, or ITS2 gene markers. The kDNA PCR and sequencing allowed the detection of L. infantum in horse blood samples. To our knowledge, this is the first report of equine infection with L. infantum in Southern Brazil. These results proved that L. infantum could also infect horses in addition to humans and dogs, as well as in European countries. This conclusion emphasizes the urgent need to follow up investigation of the infection in these animals.

The aim of this study was to verify the feasibility of synovial fluid (SF) analysis for the parasitological diagnosis of leishmaniasis in naturally infected dogs. Aseptic arthrocentesis of the tarsal and carpal joints of 40 infected dogs (158 Leishmania sp samples) was conducted after euthanasia. The SF smears were evaluated under optic microscopy and amastigotes were detected in the SF of 35 dogs (87.5%). Of the 93 positive samples, 53% were obtained from the tarsal joints, and 47% were obtained from the carpal joints. Amastigote forms were confirmed in one (17%), two (26%), three (31%) or all four investigated joints (26%). This research highlights the feasibility and high sensitivity of SF evaluation for the parasitological diagnosis of leishmaniasis in symptomatic dogs.

The relationship of animals and humans has begun many centuries ago and highlights the interdependence of humankind and some animal species. This proximity leaded to a large amount of stray dogs, cats and horses in urban centers, mining the basic precepts of welfare and causing public health problems. It can be worse in cities where concepts of animal welfare and zoonosis are not known by the population. Uruguaiana faces a critical public health situation due to the huge number of stray animals. “Best Friend” Project was created in 2010 and since then aimed to promote responsible ownership of animals, focusing on children aged 5 to 8 years old. The project worked with a theater of puppets and the performances were made in nine public schools, for 930 children. Multiple choice questions were answered by the children before and after the acting performances, in order to access their learning. The results were tabulated and analyzed by using Proportions Test (95% probability). It was observed that 89.5% of children owned pets. Before the presentation, 80% answered that the best place to keep their pet was at home, after the presentation this proportion increased to 91% (p=0.007). When inquired about the ideal food to pets, only 89% referred to commercial feed; after the project visit, the proportion increased to 99.5% (p=0.000). When asked if they would look for professional help if their pet was sick, before and after the presentation, children answered similarly (81.5% answered positively before and 80.5% after) not revealing statistical difference. Around 62% of the children considered their pet as a best friend, after the lectures 87.5% of them recognize their pet as a friend (p=0.001). It was also inquired if the children washed their hands after playing with their pet; before the educational activities of Best Friend Project 97% answered positively; after them, 100% (p=0.023). According to the evaluations, the method applied proved to be effective, improving children’s way of thinking concerning their pet maintenance. The authors believe that “Best Friend” project actions would be useful on animal welfare and public health improvements in Uruguaiana in the future.

The purpose of the present study was to compare the susceptibility to four antifungal agents of 69 Cryptococcus neoformans strains isolated from AIDS patients with that of 13 C. neoformans strains isolated from the environment. Based on the NCCLS M27-A methodology the Minimal Inhibitory Concentrations (MICs) obtained for amphotericin B, itraconazole and ketoconazole were very similar for clinical and environmental isolates. Clinical isolates were less susceptible to fluconazole than environmental isolates. The significance of these findings and aspects concerning the importance, role and difficulties of C. neoformans susceptibility testing are also discussed.