Traumatic chylothorax in urbanized free-ranging black-tufted marmosets (Callithrix penicillata).

After fatal traumatic injuries, three urbanized free-ranging marmosets developed a milky white or pink-white thoracic alkaline effusion with high specific gravity, triglyceride levels, and predominance of small lymphocytes. Chylothorax is an uncommon thoracic fluid accumulation in animals and humans and has not been reported in free-ranging non-human primates.

New insights on the phylogeography of Hepatozoon canis in Brazil.

This study aimed to molecularly characterize the Hepatozoon spp. infecting domestic and wild dogs in Brazil. A total of 22 whole blood samples tested positive for Hepatozoon spp., and five samples were sequenced for the 18S rDNA gene from H. canis after PCR amplification with four primer sets. Phylogenetic analysis using Bayesian inference showed that the three H. canis isolates from domestic dogs were not monophyletic; however, they were more closely related to each other than to other H. canis sequences. The isolate from the hoary fox (Lycalopex vetulus) was phylogenetically more distant. Two haplotype networks were constructed, identifying 10 haplotypes of H. canis in Brazil, with H10 constituting the largest group. It contains nine isolates, including three from domestic dogs. The H5 haplotype grouped the sequence of L. vetulus with two additional sequences from hosts Tapirus terrestris and L. vetulus, representing the sole haplotype with wild hosts. Bayesian analysis suggested the possible existence of two genetic groups of H. canis in Brazil, indicating gene flow of this agent within the country. These findings contribute valuable insights for a more comprehensive understanding of the molecular diversity of Hepatozoon spp. in Brazil and may help in the development of effective control measures.

Piroplasmid infection is not associated with clinicopathological and laboratory abnormalities in cats from Midwestern Brazil.

Feline piroplasmids include the genera Babesia spp., Cytauxzoon spp., and Theileria spp. In Brazil, there are few reports regarding these hemoprotozoans; however, clinicopathological and molecular data are scarce. This study aimed to characterize the clinical relevance of these parasites through hematological, biochemical, and molecular approaches. For this purpose, 166 cats from Brasilia, Federal District, Midwestern Brazil, were screened using a quantitative polymerase chain reaction (qPCR) for piroplasmids based on the LSU4 mitochondrial gene, which resulted in an overall prevalence of 36/166 (21.7%). Twelve of 166 samples (7.2%) were positive for C. felis, while 19/166 (11.4%) were positive for Babesia vogeli. No samples tested positive for Theileria spp. Babesia vogeli and Cytauxzoon spp. LSU4 sequences showed identities of 97-100% and 99.3%, respectively, to US isolates. The hematological and biochemical findings did not differ significantly between the cats that tested positive and negative for piroplasmids. Although the lack of abnormalities in clinical and laboratory parameters does not eliminate the possibility that these cats were sick and recovered, it may suggest that the Brazilian strain of Cytauxzoon spp. is not as pathogenic as that from the USA, despite the high molecular identity with North American isolates.

Eurytrema coelomaticum natural infection in small ruminants: a neglected condition.

Pancreatic eurytrematosis (PE) is an under diagnosed and neglected parasitosis in goats and sheep in the Americas. Clinical and pathological features of PE are not well defined in small ruminants worldwide. Natural cases of PE in small ruminants were detected in the Federal District, Brazil. A survey of necropsy records, including epidemiological and clinicopathological data, in goats and sheep was conducted. Most cases of PE occurred during the rainy season in adult females, with an incidence of 12.9% in goats and 0.8% in sheep. Clinical signs varied from asymptomatic infections to anorexia, lethargy, weakness, marked weight loss and death in some goats. Overall, most cases of PE in goats and sheep were incidental necropsy findings with minor pancreatic lesions. Three goats, however, showed severe chronic pancreatitis, dilation of major pancreatic ducts with numerous trematodes present and marked abdominal fat necrosis. Morphological and molecular characterization of flukes detected Eurytrema coelomaticum. Our findings shed light on the prevalence of E. coelomaticum infections in small ruminants in the region and highlight the possibility of severe and lethal cases in goats. PE must be further investigated in small ruminant populations in relevant livestock production regions of the Americas.

Canine piroplasmids: Molecular detection and laboratory characterization in dogs from Brasilia, Brazil, with the first molecular evidence of dog exposure to a novel opossum-associated Babesia sp.

Canine piroplasmid infections can be caused by Babesia spp., Theileria spp. and Rangelia vitalii. In Brazil, canine babesiosis caused by Babesia vogeli is endemic and reported throughout the country. On the other hand, Rangeliosis caused by R. vitalii has only been described so far in the South and Southeast regions. Despite that, studies analyzing the laboratory and molecular characterization of these hemoprotozoa are still scarce. To investigate the occurrence, the laboratory features, the molecular characterization, and the diversity of piroplasmids from Midwestern Brazil, a survey was performed using blood samples obtained from 276 domestic dogs from Brasília, Federal District, Midwestern Brazil. A broad-range quantitative PCR (qPCR) targeting the mitochondrial large subunit ribosomal DNA (LSU4) was used to detect piroplasmid DNA. The overall molecular occurrence of piroplasmids was 11.2% (31/276), with 9.7% (27/276) of the sequences identified as Babesia vogeli (98-100% identity to B. vogeli isolate from the USA). Based on a partial 18S rRNA sequence pairwise alignment (-250 bp), 1.4% (4/276) of the sequences showed only 76.8% identity with B. vogeli but 100% identity with opossum-associated Babesia sp. (MW290046-53). These findings suggest the exposure of dogs from Brazil to a recently described Babesia sp. isolated from white-eared opossum. None of the analyzed dogs was positive for Theileria spp. or R. vitalii. Subsequently, all positive sequences were submitted to three additional PCR assays based on the 18S rRNA, cox-1, and cytb genes, aiming at performing a haplotype network analysis. Haplotype network using cox-1 sequences showed the presence of six different haplotypes of B. vogeli; one of them was shared with isolates from Brazil, the USA, and India. When including animals co-infected with other vector-borne diseases, piroplasmid-positive dogs had 2.3 times higher chance of having thrombocytopenia than the negative ones. The molecular results demonstrated that the compared Babesia vogeli sequences showed a low variability as well as evidence of exposure to a putative novel opossum-associated Babesia sp. in dogs from Midwestern Brazil.

Molecular detection of Leucocytozoon in red-legged seriemas (Cariama cristata), a non-migratory bird species in the Brazilian Cerrado.

Avian Haemosporidian parasites - Plasmodium, Haemoproteus, Leucocytozoon and Fallisia - have a wide distribution except for Antarctica. Leucocytozoon sp. has been poorly described in Brazil, and few studies have indicated infections in birds from the Atlantic Forest, Pantanal, Pampa and Amazon biomes. This study describes, for the first time, the occurrence of Leucocytozoon infection in red-legged seriemas (Cariama cristata) in the Brazilian savanna (Cerrado biome) using molecular diagnosis. Leucocytozoon spp. lineage CARCRI01 was detected in three C. cristata, a non-migratory bird, confirming transmission in mid-elevation areas in central Brazil. Further studies are needed to certify whether infections in red-legged seriemas were not abortive and to elucidate Leucocytozoon infection at low altitudes in the Brazilian lands.

miR-20a is upregulated in serum from domestic feline with PKD1 mutation.

Polycystic kidney disease (PKD), also known as autosomal dominant polycystic kidney disease (ADPKD) is a genetically heterogeneous condition characterized by cysts in renal parenchyma. It is the most prevalent inherited disease of domestic cats. MicroRNAs (miRNAs or ncRNA) are short, noncoding, single-stranded RNAs that may induce PKD cytogenesis by affecting numerous targets genes as well as by directly regulating PKD gene expression. We compared the relative expression profile of miR-20a, -192, -365, -15b-5p, and -16-5p from plasma and serum samples of nine domestic cats with PKD1 mutation, detected by polymerase chain reaction (PCR), and a control group (n = 10). Blood samples from cats with PKD1 mutation provide similar concentrations of microRNAs either from plasma or serum. Serum miR-20a is upregulated in PKD group with p < 0.005; Roc curve analysis showed an AUC of 90,1% with a cut-off value sensitivity of 77.8% and specificity of 100%. This data provides important information regarding renal miRNA expression in peripheral blood sampling.

Insights from the use of erythropoietin in experimental Chagas disease.

In addition to the long-established role in erythropoiesis, erythropoietin (Epo) has protective functions in a variety of tissues, including the heart. This is the most affected organ in chronic Chagas disease, caused by the protozoan Trypanosoma cruzi. Despite seven million people being infected with T. cruzi worldwide, there is no effective treatment preventing the disease progression to the chronic phase when the pathological involvement of the heart is often observed. Chronic chagasic cardiomyopathy has a wide variety of manifestations, like left ventricular systolic dysfunction, dilated cardiomyopathy, and heart failure. Since Epo may help maintain cardiac function by reducing myocardial necrosis, inflammation, and fibrosis, this study aimed to evaluate whether the Epo has positive effects on experimental Chagas disease. For that, we assessed the earlier (acute phase) and also the later (chronic phase) use of Epo in infected C57BL/6 mice. Blood cell count, biochemical parameters, parasitic load, and echocardiography data were evaluated. In addition, histopathological analysis was carried out. Our data showed that Epo had no trypanocide effect nor did it modify the production of anti-T. cruzi antibodies. Epo-treated groups exhibited parasitic burden much lower in the heart compared to blood. No pattern of hematological changes was observed combining infection with treatment with Epo. Chronic Epo administration reduced CK-MB serum activity from d0 to d180, irrespectively of T. cruzi infection. Likewise, echocardiography and histological results indicate that Epo treatment is more effective in the chronic phase of experimental Chagas disease. Since treatment is one of the greatest challenges of Chagas disease, alternative therapies should be investigated, including Epo combined with benznidazole.

Use of multivariate analyses for determining heat tolerance in Brazilian cattle.

Adaptability can be evaluated by the ability of an animal to adjust to environmental conditions and is especially important in extreme weather conditions such as that found in tropical Brazil. A multivariate analysis using physical and physiological traits in exotic (Nellore and Holstein) and naturalized (Junqueira, Curraleira, Mocho Nacional, Crioula Lageana, and Pantaneira) cattle breeds was carried out in the Federal District of Brazil to test and determine which traits are important in the adaptation of animal to heat stress as well as the ability of these traits and statistical techniques to separate the breeds studied. Both physical and physiological traits were measured on three occasions and included body measurements, skin and hair thickness, hair number and length, pigmentation, sweat gland area as well as heart and breathing rates, rectal temperature, sweating rate, and blood parameters. The data underwent multivariate statistical analyses, including cluster, discriminate, and canonical procedures. The tree diagram showed clear distances between the groups studied, and canonical analysis was able to separate individuals in groups. Coat traits explained little variation in physiological parameters. The traits which had higher discriminatory power included packed cell volume, shoulder height, mean corpuscular volume, body length, and heart girth. Morphological and physiological traits were able to discriminate between the breeds tested, with blood and size traits being the most important. More than 80% of animals of all breeds were correctly classified in their genetic group.

Hematological and Serum Biochemical Changes and Their Prognostic Value in Horses Spontaneously Poisoned by Crotalaria spectabilis.

Determining the prognosis of poisoning by plants containing pyrrolizidine alkaloids is usually challenging. This study aimed to identify important prognostic parameters that can determine the severity of spontaneous poisoning by Crotalaria spectabilis in horses. Blood samples from 42 horses spontaneously poisoned by oats contaminated with C. spectabilis seeds were evaluated. Complete blood counts (CBC) and serum biochemical tests [urea, creatinine, total protein, albumin, total and direct bilirubin concentrations, aspartate aminotransferase (AST), γ-glutamyl transferase (GGT), and creatine kinase (CK) activities] were performed. Horses were followed up for 12 months to determine the long-term survival rate; after 12 months, they were divided into two groups: survivors (n = 30) and non-survivors (n = 12). Horses spontaneously poisoned with C. spectabilis had higher levels of urea, globulin, bilirubin (total, direct, and indirect), AST, GGT, and CK than the reference values. Non-survivor horses showed significantly higher (p < 0.05) values of hemoglobin, GGT, and direct bilirubin than the survivor horses. Horses with serum GGT activity higher than 95 U/l had 14.0 times the risk of death compared to animals showing activities equal to or lower than this value, whereas horses with serum direct bilirubin concentration higher than 0.6 mg/dl (10.26 µmol/L) had 5.78 times the risk of death compared to the others. In summary, serum GGT activity and direct bilirubin concentration may be useful prognostic indicators for assessing the severity of C. spectabilis-poisoned horses.

Capybara (Hydrochoerus hydrochaeris) exposure to Rickettsia in the Federal District of Brazil, a non-endemic area for Brazilian spotted fever.

In this study, whole blood samples and ticks were collected from 57 capybaras in recreational areas in the Federal District, Brazil, aiming to investigate the presence of Rickettsia spp. using polymerase chain reaction (PCR) and indirect immunofluorescence (IFAT) assays. None of the capybara blood samples yielded rickettsial DNA by PCR. Among 55 capybara serum samples tested by IFAT, 53 (96.3%) reacted to Rickettsia spp. Among these, 21 (39.6%) identified the R. bellii antigen as the probable antigen involved in a homologous reaction (PAIHR), whereas 2 (3.8%) identified the R. parkeri antigen. Ticks collected from capybaras were identified as 173 Amblyomma sculptum and 410 A. dubitatum, in addition to nine Amblyomma spp. larvae. A sample of 231 ticks was subjected to DNA extraction and PCR for Rickettsia species. None of 122 A. sculptum yielded rickettsial DNA. Molecular evidence of R. bellii was found in 25/108 (23.1%) and of Rickettsia sp. strain Cooperi (R. parkeri-like agent) in 2/108 (1.9%) of the A. dubitatum samples. These results suggest a greater exposure to R. bellii in these capybara populations, in addition to a more significant number of A. dubitatum, which might characterize the Federal District region as not endemic for Brazilian spotted fever.

Multivariate analysis for characteristics of heat tolerance in horses in Brazil.

The environment in which the horse is reared affects its ability to maintain thermal balance which is in turn related to thermal characteristics and regulatory physiological mechanisms. In this study a multivariate analysis of physiological traits in relation to heat tolerance in horses was carried out in the Federal District, Brazil. The aim was to test the ability of these analyses to separate groups of animals and determine which physiological traits are most important in the adaptation to heat stress. Forty adult horses (4 to 13 years) were used, ten from each of four different genetic groups (English thoroughbred, Brazilian showjumper, crossbred and Breton). The traits examined included heart and breathing rate, rectal temperature as well as blood parameters. The data underwent multivariate statistical analysis including cluster, discriminate and canonical using Statistical Analysis System - SAS (R) procedures CLUSTER, STEPDISC, CANCORR and DISCRIM. The tree diagram showed clear distances between groups studied and canonical analysis was able to separate individuals in groups. The discriminate analysis identified the variables which were most important in separating these groups. The multivariate analysis was able to separate the animals into groups with RR, HR and RT being important in this separation.

Canine atopic dermatitis attenuated by mesenchymal stem cells.

OBJECTIVE: To evaluate the use of mesenchymal stem cells (MSCs) in the attenuation of canine atopic dermatitis (AD). MATERIALS AND METHODS: Sixteen dogs were selected and divided into three groups, mild, moderate, and severe, according to the Canine Atopic Dermatitis Extent and Severity Index (CADESI-4). They were evaluated for 82 days. The protocol recommended in this experiment was to inject 2 × 106/kg bodyweight of MSC's in all groups by the intravenous route with intervals of applications of 21 days. The degree of pruritus was evaluated by examining the visual analog scale, the CADESI-4, the histopathology of the skin, hematological and biochemical parameters, the pyogenic effect of MSCs, and the thickness of the epidermis. RESULTS: There was a significant difference in the reduction of epidermal thickness in the moderate and severe groups. Hematological, biochemical, and body temperature parameters remained within normal limits for the species with no side effects. CONCLUSION: MSCs attenuated the clinical signs of AD.

Fetal development and blood hematological-biochemical parameters in Campeiro and Pantaneiro foals.

For a long time, Pantaneiro and Campeiro breeds were raised only within their places of origin. Consequently, there are few of these horses; therefore, establishing reproductive and clinical standards for these animals is necessary to implant new biotechnologies for reproduction to preserve their genetics. This study aimed to perform a descriptive evaluation of fetal age determination by fetus ocular orbit measurement in mares of the Campeiro and Pantaneiro breeds. We also evaluated sequential changes in hematology and biochemistry for foals from birth to six months of life by counting red blood cells, hemoglobin concentration, mean corpuscular volume, mean corpuscular hemoglobin concentration, leukocytes, neutrophils, lymphocytes, eosinophils, monocytes, basophils, platelet count, fibrinogen, albumin, plasma protein, creatinine, aspartate aminotransferase, alkaline phosphatase, gamma-glutamyl transferase, and globulins. There was no significant difference in relation to fetal gestational age and ocular orbit between the two breeds (P = 0.578). There was no significant difference in the hematological parameters between the Campeiro and Pantaneiro foals, but there were differences in the means and changes in the blood variables when compared to the literature. These hematological and biochemical variations provide useful information for clinical evaluations from Campeiro and Pantaneiro foals up to six months of age.

Theileria equi infection causing abortion in a mare in Brazil.

The aim of this report was to confirm the cause of abortion to be Theileria equi acquired transplacentally in a mare in Federal District, Brazil. The aborted fetus showed a high parasitemia of erythrocytes (80%) by the oval and Maltese cross forms of T. equi. Necropsy of the fetus demonstrated congestion of organs, jaundice, anemia and pronounced spleno- and hepatomegaly. Twelve breeding mares, including the one that aborted and her fetus, tested positive for T. equi infection by universal and multiplex PCR assay. The absence of histopathologic changes such as inflammation and necrosis in the placenta and fetal tissues as well as the negative microbiological results and negative serology for Leptospira spp. in the mare that aborted were important to exclude other infectious agents that may be involved in equine abortion. This report highlights the transplacental transmission of T. equi infection in horses in endemic areas and reinforces the importance of investigating equine theileriosis as a possible cause of abortion in mares.

Clinical, hematological, biochemical, and ultrasonographic aspects of Platynosomum sp. (Trematoda: Dicrocoeliidae) infection of captive Callithrix penicillata.

Trematodes from the genus Platynosomum have been found to infect Neotropical primates in captivity, but little is known about their pathogeny in such hosts. This study evaluated the physiological effects of natural infection by the liver-dwelling trematode Platynosomum sp. in ten males and ten females of Callithrix penicillata kept in captivity at the Primate Center of the University of Brasília. The marmosets were examined twice, 6 months apart. The following parameters were analyzed: complete blood count, bleeding time, serum total protein, albumin, and the liver enzymes AST and ALT, and both a stool analysis and a liver ultrasonic evaluation were performed. We were able to characterize a group of abnormalities associated with this trematode infection which were mainly derived from the hepatitis caused by it: coagulation disorders, abnormal red blood cells, hypoalbuminemia, and abnormal levels of liver-linked serum enzymes. Eosinophilia and thrombocytopenia were also commonly seen. All of the aforementioned abnormalities were in good accord with typical effects of trematodes on liver parenchyma. We suggest that this set of abnormalities is characteristic of the infection of C. penicillata with Platynosomum sp., and should be among the most prominent aspects that the veterinary surgeon considers when suspecting such an infection. We also suggest that these clinical signs and abnormalities will be similar in other liver-dwelling trematode-infected primate species.

Hepatozoon spp.: report of some cases in dogs in Brasília, Brazil.

Canine hepatozoonosis is a disease caused by the tick-borne protozoan Hepatozoon spp. It has been reported in the United States, southern Europe, the Middle East, Africa and the Far East. In Brazil, canine hepatozoonosis is an emerging protozoal tick-borne disease, and is characterized by distinct clinical signs. The objective of this study was to analyze the laboratory findings of some hepatozoonosis cases in dogs in Brasília, Brazil, and their clinical signs. The animals of this experiment showed low parasitemia, similar to H. americanum, but the clinical signs presented were similar to H. canis. According to our observations and in agreement with O'Dwyer et al. [Vet. Parasitol. 94 (2001) 143], the Brazilian Hepatozoon appears more to resemble the species found in the eastern Hemisphere than with H. americanum of North America, or could be caused by a new species. Our data revealed that hepatozoonosis could be considered endemic in Brasília.

Prevalence and phylogenetic analysis of haemoplasmas from cats infected with multiple species.

Mycoplasma haemofelis (Mhf), 'Candidatus Mycoplasma haemominutum' (CMhm) and 'Candidatus Mycoplasma turicensis' (CMt) are agents of feline haemoplasmosis and can induce anaemia in cats. This study aimed to determine the prevalence and phylogeny of haemoplasma species in cats from Brazil's capital and surrounding areas, and whether correlation with haematological abnormalities existed. Feline haemoplasmas were found in 13.8% of 432 cats. CMhm was the most prevalent species (in 13.8% of cats), followed by Mhf (11.1%) and CMt (4.4%). Over 80% of haemoplasma-infected cats harboured two or more feline haemoplasma species: 7.1% of cats were co-infected with Mhf/CMhm, 0.4% with CMhm/CMt and 3.9% with Mhf/CMhm/CMt. Male gender was significantly associated with haemoplasma infections. No association was found between qPCR haemoplasma status and haematological variables, however CMhm relative copy numbers were correlated with red blood cell (RBC) numbers and packed cell volume (PCV). Haemoplasma 16S rRNA gene sequences (> 1 Kb) were derived from co-infected cats using novel haemoplasma species-specific primers. This allowed 16S rRNA gene sequences to be obtained despite the high level of co-infection, which precluded the use of universal 16S rRNA gene primers. Within each species, the Mhf, CMhm and CMt sequences showed > 99.8%, > 98.5% and > 98.8% identity, respectively. The Mhf, CMhm and CMt sequences showed > 99.2%, > 98.4% and > 97.8% identity, respectively, with GenBank sequences. Phylogenetic analysis showed all Mhf sequences to reside in a single clade, whereas the CMhm and CMt sequences each grouped into three distinct subclades. These phylogeny findings suggest the existence of different CMhm and CMt strains.

Detection and molecular characterization of piroplasms species from naturally infected dogs in southeast Brazil.

Rangelia vitalii is a protozoon described from dogs in the south and southeast regions of Brazil. It is phylogenetically related to Babesia spp. that infects dogs, but data on this enigmatic parasite is still limited. The aim of this work was to detect piroplasm species in dogs in the state of Rio de Janeiro, Brazil, by 18S rRNA gene-based PCR assay, restriction fragment length polymorphism (RFLP) and sequence analyses. Of 103 dogs examined, seven (6.8%) were positive for Babesia spp. by PCR. The amplified products were digested by restriction enzymes to differentiate the Babesia species, and one sample was identified as Babesia vogeli. The pattern observed for the other six amplification products did not match with pattern described for large Babesia infecting dogs. Sequencing analysis confirmed these six samples as R. vitalii, with high homologies (99-100%) with a sequence from south Brazil. This study confirms the presence of Babesia vogeli and Rangelia vitalii circulate in domestic dogs in Teresópolis, Rio de Janeiro, Brazil.

Heat tolerance in Brazilian sheep: physiological and blood parameters.

Thirty Santa Ines adult, non-lactating, non-pregnant ewes, ten with a brown coat, ten black coated and ten white coated, as well as ten Bergamasca and ten of mixed breed were used to evaluate the effect of climate on physiological and blood parameters in sheep. Two sample collections were taken (6 AM: and 2 PM: ) on six days. Sweating rate (SR), heart (HR) and breathing rates (BR), complete hemogram, rectal (RT) and skin temperatures (ST) were measured. Variance analyses were carried out using SAS(R). In general, there were significant differences between animals due to skin type, time and day of collection. White coated animals showed lower HR, BR and RT, with afternoon parameters higher than morning. Correlations between HR, BR, RT, SR and ST were medium and positive. Correlations between physiological and blood traits were in general negative and medium. Packed Cell Volume (PCV), total plasma proteins, red blood cell count and hemoglobin concentration had high positive correlations between each other. The first two autovectors explained 49% of variation between traits. White coated Santa Ines animals were shown to be better adapted to climatic conditions in Central Brazil and wool sheep more affected by heat.