Biomarkers of gastrointestinal nematodes in beef cattle raised in a tropical area.

Biomarkers are specific molecular, histological, or physiological characteristics of normal or pathogenic biological processes and are promising in the diagnosis of gastrointestinal nematodes (GINs). Although some biomarkers have been validated for infection by Ostertagia sp. in cattle raised in temperate regions, there is a lack of information for tropical regions. The aim of this project was to assess potential biomarkers and validate the most promising. In the first study, 36 bovines (Nelore breed) naturally infected by GINs were distributed into two groups: infected (not treated with anthelmintic) and treated (treated with fenbendazole on days 0, 7, 14, 21, 28, 42, and 56). The variables of interest were live weight, fecal egg count, hemogram, serum biochemical markers, phosphorus, gastrin, and pepsinogen. In the second step, pepsinogen was assessed in cattle of the Nelore breed distributed among three groups: infected (not treated with anthelmintic), MOX (treated with moxidectin), and IVM + BZD (treated with ivermectin + albendazole). In the first study, no difference between groups was found for weight, albumin, hematocrit (corpuscular volume [CV]), erythrocytes, or hemoglobin. Negative correlations were found between pepsinogen and both CV and albumin, and albumin was negatively correlated with the percentage of Haemonchus sp. in the fecal culture. Among the biomarkers, only pepsinogen differentiated treated and infected (beginning with the 28th day of the study). In the second study, a reduction in pepsinogen was found after anthelmintic treatment. Therefore, pepsinogen is a promising biomarker of worms in cattle naturally infected by the genera Haemonchus and Cooperia in tropical areas.

Turning chaotic sample group clusterization into organized ones by feature selection: Application on photodiagnosis of Brucella abortus serological test.

Bovine brucellosis diagnosis is a major problem to be solved; the disease has a tremendous economic impact with significant losses in meat and dairy products, besides the fact that it can be transmitted to humans. The sanitary measures instituted in Brazil are based on disease control through diagnosis, animal sacrifice, and vaccination. Although the currently available diagnostic tests show suitable quality parameters, they are time-consuming, and the incidence of false-positive and/or false-negative results is still observed, hindering effective disease control. The development of a low-cost, fast, and accurate brucellosis diagnosis test remains a need for proper sanitary measures at a large-scale analysis. In this context, spectroscopy techniques associated with machine learning tools have shown great potential for use in diagnostic tests. In this study, bovine blood serum was investigated by UV-vis spectroscopy and machine learning algorithms to build a prediction model for Brucella abortus diagnosis. Here we first pre-treated the UV raw data by using Standard Normal Deviate method to remove baseline deviation, then apply principal component analysis - a clustering method - to observe the group formation tendency; the first results showed no clustering tendency with a messy sample score distribution, then we properly select the main principal components to improve clusterization. Finally, by using machine learning algorithms (SVM and KNN), the predicting models achieved a 92.5% overall accuracy. The present methodology provides a test result in an average time of 5 min, while the standard diagnosis, with the screening and confirmatory tests, can take up to 48 h. The present result demonstrates the method's viability for diagnosing bovine brucellosis, which can significantly contribute to disease control programs in Brazil and other countries.

Canine visceral leishmaniasis diagnosis by UV spectroscopy of blood serum and machine learning algorithms.

Visceral leishmaniasis (VL) is a zoonotic disease caused by the protozoan Leishmania infantum, and dogs are considered the main urban hosts for future disease transmission. The first and most effective control against the spread of disease relies on identifying infected animals, followed by their treatment or sacrifice, to reduce the protozoan reservoirs. Despite the availability of various diagnostic tests for VL in dogs the development of a quick and accurate diagnosis is essential from a public health and ethical point of view. Here we analyze the use of UV-Vis spectroscopy as an alternative diagnostic method for VL diagnosis by using the antigen-antibody interaction in canine blood serum and machine learning algorithms. The main UV spectra in the 220 to 280 nm range exhibit nine electronic absorption bands, but no significative difference could be identified between the positive and negative group spectra. Finally, UV pre-proceed spectra by SNV (standard normal variate) were submitted to principal component analysis followed by Linear SVM algorithm, the prediction model was tested in a leave-one-out cross-validation and external validation test reaching an overall accuracy of 75%.

Molecular characterization of Trypanosoma cruzi DTUs of the triatomine species in a Chagas disease endemic area.

Trypanosoma cruzi is the etiological agent of Chagas disease, a neglected tropical infection with great public health importance. This protozoan has triatomine insects as vector but may also be transmitted through blood transfusion, organ transplants, ingestion of contaminated food, or congenitally. It has a heterogeneous population classified into Discrete Typing Units (DTUs), TcI-TcVI and TcBat. The aim of this study was to molecularly characterize the DTUs of T. cruzi in triatomines from a Chagas disease endemic area in Northeastern Brazil. Triatomines were collected and the gut content was microscopically analyzed to investigate the presence of trypanosomatid flagellates. In addition, digestive tracts of some specimens were dissected and molecularly analyzed through PCR for Trypanosoma spp. and sequencing. PCR positive samples were further submitted to a multiplex PCR for DTUs of T. cruzi. A total of 117 triatomines were collected, 93.16% being in intradomicile and 6.84% in peridomicile environments. Insects were identified as Panstrongylus lutzi (37.60%), Triatoma pseudomaculata (26.50%), Triatoma brasiliensis (23.08%) and Panstrongylus megistus (12.82%). The specimens herein analyzed presented infection rates by T. cruzi of 5.49% and 12.09% in parasitological and molecular examinations, respectively. Multiplex PCR screening revealed 70.59% of the TcI genotype, detected in all triatomine species identified in this study and 29.41% of the DTU TcIII/TcIV detected in P. megistus and P. lutzi. T. cruzi infect triatomines in intradomicile and peridomicile environments, which brings attention to the risk of human infections and to the importance of the implementation of surveillance and entomological control actions.

SALMONELLA SPP. INFECTION IN PSITTACIDAE AT A WILDLIFE REHABILITATION CENTER IN THE STATE OF MATO GROSSO DO SUL, BRAZIL.

The growing interest in wild birds as pets and the increasingly complex interspecific relationships due to human activities in wild environments underscore the need for better knowledge about the health of these animals. Salmonella stands out among the infectious agents of considerable importance to both animal and human health. The importance of these enterobacteria to the health of humans and livestock animals has long been known. In wild birds in countries such as Brazil, however, little is known regarding the frequency of infection and the main serotypes of occurrence. In the present study, the frequency of infection and the main serotypes of Salmonella spp. were investigated in 258 birds at a wild animal rehabilitation center using conventional microbiological methods and molecular diagnostic techniques. Four birds infected with Salmonella enterica were identified using PCR. The birds were of the species Brotogeris chiriri, Ara ararauna, and Eupsittula aurea. Sequencing of DNA revealed identity with the Javiana, Newport, and Arizonae serotypes. These results are of considerable importance to the implementation of management and control measures directed at human and animal health.

Biomarkers and renal arterial resistive index in dogs naturally infected with Leishmania infantum.

Canine visceral leishmaniasis frequently causes glomerulonephritis and tubulointerstitial nephritis, nephropathies for which diagnosis has been limited by the low sensitivity of traditional tests. The aim of this study was to evaluate serum cystatin C and urinary gamma-glutamyltransferase (uGGT) levels and the urinary GGT/urinary creatinine ratio (uGGT/uCR) and to measure the renal arterial resistive index (RARI) in dogs with leishmaniasis with varying degrees of renal injury based on the urine protein: creatinine ratio (UP/C) and serum creatinine (SCr) level. We tested 59 untreated adult dogs of both sexes and undefined breeds naturally infected with Leishmania infantum. The dogs were grouped into four groups based on UP/C and SCr level: group 1 (n = 15), dogs with SCr levels < 1.4 mg/dL and UP/C < 0.5; group 2 (n = 13), dogs with SCr levels < 1.4 mg/dL and UP/C of 0.5-1.0; group 3 (n = 16), dogs with SCr levels < 1.4 and UP/C > 1.0; and group 4 (n = 15), dogs with SCr levels > 1.4. A fifth group of healthy dogs (n = 10) was the control. uGGT concentrations and uGGT/uCR were higher in dogs with proteinuria and SCr < 1.4 mg/dL, whereas the serum cystatin C concentrations and RARI were higher only in dogs with SCr levels > 1.4. In conclusion, uGGT and uGGT/uCR may be useful tools for early detection and assessment of renal lesions associated with leishmaniasis; however, cystatin C is useful for monitoring the progression of kidney disease when measured sequentially.

Cercopithifilaria bainae in Rhipicephalus sanguineus sensu lato ticks from dogs in Brazil.

Rhipicephalus sanguineus sensu lato (s.l.) ticks act as intermediate host for a range of canine vector-borne pathogens, including nematodes ranked in the genus Cercopithifilaria. Though being the object of several studies in the last years, information on the distribution of these parasites is still lacking. In this study, the occurrence of Cercopithifilaria spp. was investigated in on-host population of R. sanguineus s.l. collected from naturally infested dogs. Ticks (n=1906, including one larva, 294 nymphs and 1611 adults) were sampled on domestic dogs (n=155) living in the municipality of Garanhuns (northeastern Brazil). Tick collections (n=36) were performed every 8 days, from October 2015 to June 2016. Filarioid larvae detected at tick dissection were morphologically and morphometrically identified at species level. At the end of the study, only R. sanguineus s.l. ticks were collected, with the highest number in January 2016 (n=254) and the lowest in June 2016 (n=26). Out of 1906 dissected ticks, 2.68% (51/1906) harboured Cercopithifilaria bainae larvae, whose identification was molecularly confirmed, with a nucleotide identity of 99% with C. bainae. Data here reported indicate that, in the study area, R. sanguineus s.l. is the predominant tick infesting domestic dogs. Accordingly, these animals are at a high risk of C. bainae infection.

Larvae of Ixodiphagus wasps (Hymenoptera: Encyrtidae) in Rhipicephalus sanguineus sensu lato ticks (Acari: Ixodidae) from Brazil.

The biological control of ticks represents an alternative method to the chemical control, given its ecological-friendly approach. Amongst the alternatives, the use of parasitoids of the genus Ixodiphagus (Hymenoptera: Encyrtidae) has been largely investigated. The aim of this study was to document and molecularly characterize Ixodiphagus wasps in ticks from a tropical region of Brazil. From October 2015 to March 2016, Rhipicephalus sanguineus sensu lato ticks (n=1814) were collected from naturally infested dogs and Ixodiphagus larvae were detected by microscopic examination. In addition, adult wasps were obtained in the laboratory. Larvae and adults were molecularly identified as Ixodiphagus hookeri. These findings suggest that this type of parasitism deserves to be studied in local tick populations, in order to elucidate the role of these wasps as a potential alternative to chemical tick control.

Natural Infection of the South American Tapir ( Tapirus terrestris ) by Theileria equi.

Theileria equi is a tick-borne piroplasm considered endemic in equines in Brazil. The cohabitation of domestic and wild animals in areas of extensive cattle breeding favors the close contact between different species and the sharing of vectors and, consequently, pathogens. We report the natural infection of a young South American tapir ( Tapirus terrestris ) by T. equi in Mato Grosso do Sul, Brazil. Although it was not possible to associate the clinical and hematologic status of the animal with the infection by the protozoan parasite, our report represents an alert on the sharing of pathogens between domestic and wild animals.

Filarioids infecting dogs in northeastern Brazil.

Species of filarial nematodes belonging to the genera Dirofilaria and Acanthocheilonema are recognised as common parasites of dogs throughout the world. Recently, other filarioids featured by the presence of dermal microfilariae (e.g., Onchocerca lupi and Cercopithifilaria spp.) have been recognised in Europe. In Brazil, reports of filarioids in dogs are limited to Dirofilaria immitis, Acanthocheilonema reconditum and Cercopithifilaria bainae. To investigate the distribution of filarial infections in dogs living in an endemic region from northeastern Brazil, blood and skin samples (n=104) were microscopically (modified Knott's test and skin snip sediment examination) and molecularly evaluated. Twenty-two dogs (21.15%) were positive at microscopic and/or molecular examination for at least one filarioid species, with 21 (20.19%) animals positive for blood microfilariae at molecular and/or at microscopic examination. Microfilariae of D. immitis were detected in 12 (11.54%) animals, with co-infection of D. immitis and A. reconditum observed in four (3.85%) individuals. One animal was positive for C. bainae at both microscopic and molecular examination. Analysis of sequence obtained in the present study showed significant alignment identity with that of C. bainae from Europe. Considering that in the area of study arthropod vectors (mosquitoes, fleas and ticks) are prevalent throughout the year, preventive measures should be disposed in order to avoid the animal infestation and pathogen infection.