Impact of blood storage duration on hematologic, blood gas, biochemical, and oxidative stress variables in sheep undergoing allogeneic blood transfusions.

BACKGROUND: Hemotherapy in ruminants is limited to whole blood transfusions, sometimes with stored blood for up to 42 days, but little attention has been given to the effect of blood storage times and recipient responses after transfusions. OBJECTIVES: We aimed to evaluate the hematologic and serum biochemical effects after allogeneic blood transfusion with either fresh or stored blood in sheep. We also sought to examine hematologic and biochemical analyte changes in the store blood. METHODS: Eighteen sheep underwent a single phlebotomy to remove 40% of their blood volume. The sheep were divided into three experimental groups, G0, G15, and G35, which included six animals, each receiving 20 mL/kg of either fresh blood or blood stored in citrate, phosphate, dextrose, and adenine (CPDA-1) bags for 15 and 35 days, respectively. Biochemical, hematologic, coagulation, blood gas, lipid peroxidation, and oxidative stress test evaluations were performed using the blood samples gathered at T0 (before transfusion), 30 minutes (T30m), 6, 12, 24, 48, 72, and 96 hours (T6h-T96h), 8 days (T8d), and 16 days (T16d) after transfusions. RESULTS: Sheep exhibited increases in packed cell volumes, red blood cell counts, and total hemoglobin concentrations at T30m (P < .05). G35 animals had greater plasma hemoglobin concentrations at T12h and decreased blood pH values at T6h, characterized by slight metabolic acidemia. Regarding oxidative stress, G35 animals had decreased catalase activities from T0 at T30m, T6h, T12h, and T24h, indicating that hemolysis had occurred, which was supported by concomitant increases in bilirubin. CONCLUSIONS: Sheep transfused with 35-day stored blood exhibited greater hematologic, blood gas, biochemical, and oxidative alterations; however, anemic animals without comorbidities effectively reversed those alterations.

Typhus Group Rickettsiosis, Brazilian Amazon.

Rickettsia rickettsii infection is the only rickettsiosis included in the list of reportable diseases in Brazil, where typhus group rickettsioses, mainly murine typhus, have been underreported. We report a case of typhus group rickettsiosis with unique ecologic particularities in a patient from the Brazilian Amazon, where, to our knowledge, rickettsioses have not been reported.

Influence of organic chromium supplementation on the performance of beef calves undergoing weaning-related stress / Influência da suplementação com cromo orgânico no desempenho de bezerros de corte submetidos ao estresse da desmama

Dietary chromium supplementation before, during, and after weaning was conducted to evaluate the hypothesis that chromium supplementation could reduce weaning-induced cortisol release in beef calves. We examined the effects of chromium supplementation in 150 crossbred calves (male and female) between five and six months of age. The calves were randomly divided by sex and breed into two equal homogeneous groups (n=75). One group was used as the control, and the other experimental group received supplementation with 0.9mg of chromium carbon-amino-phospho-chelate per 100kg BW. The chromium supplement was mixed with mineral salt for the consumption of 0.1% of BW, and the supplement was administered via creep feeding 60 days before and 60 days after forced weaning. Calves were weighed, and their blood and urine samples were obtained at four time-points: T0 (60 days before weaning), T1 (at weaning), T2 (48 hours after weaning), and T3 (60 days after weaning). Blood samples were used to determine chromium, cortisol, total protein, and albumin concentrations, and urine samples were used to determine urinary creatinine and chromium levels. Cumulative weight gain was higher in calves supplemented with chromium before weaning and during the experiment (P<0.05). In addition, weaning-related stress caused an increase in chromium excretion in the urine, and chromium supplementation reduced stress, which resulted in lower cortisol and total protein levels during weaning.(AU)

O estudo foi realizado para avaliar a hipótese de que a suplementação dietética com cromo antes, durante e após a desmama possa diminuir a concentração de cortisol causado por este processo em bezerros de corte. Para tal, foram utilizados 150 bezerros mestiços, machos e fêmeas, entre cinco e seis meses de idade. Os animais foram divididos randomicamente por sexo e grupo genético em dois grupos homogêneos (n=75), um mantido como controle e outro suplementado com 0,9mg de carboaminofosfoquelato de cromo/100 kg PV misturado a um sal proteinado para ser consumido na base de 0,1% do PV via creep feeding, no decorrer de 60 dias antes e 60 dias após à desmama forçada. Os animais foram pesados e foram coletadas amostras sanguíneas e urinárias no M0 (60 dias antes da desmama), M1 (desmama), M2 (48 horas após a desmama) e M3 (60 dias após à desmama) para determinação de cromo, cortisol, proteína total e albumina no sangue e da concentração urinária de creatinina e cromo. O ganho acumulado de peso foi superior nos bezerros suplementados com cromo antes da desmama e no decorrer de todo o experimento (P<0,05). A suplementação com cromo reduziu os teores de cortisol e de proteína total durante a desmama. O estresse da desmama provocou aumento da excreção de cromo pela urina.(AU)

Biochemical responses, blood gas, oxidative stress and lipid peroxidation of goats transfused with fresh or stored whole homologous blood / Respostas bioquímicas, hemogasométricas, estresse oxidativo e peroxidação lipídica de caprinos transfundidos com sangue homólogo total fresco ou armazenado

Blood transfusion is a therapeutic procedure of great importance for veterinary medicine, in spite of only few studies in the literature on hemotherapy in goats. We aimed to evaluate the biochemical, blood gas, oxidative stress and lipid peroxidation of goats submitted to homologous transfusion of fresh whole blood or stored for 15 and 35 days. Eighteen adult male goats were submitted to a single phlebotomy to remove 30% of the blood volume, and we transfused 20mL/kg of whole blood stored in CPDA-1 bags according to the experimental group, being: G0 composed goats who received fresh blood, G15 and G35 goats that received blood stored for 15 and 35 days, respectively. For the biochemical evaluation, blood gas, oxidative stress and lipid peroxidation, blood samples were collected at the following moments: before the induction of anemia (TC0); 6 hours after phlebotomy and before transfusion (TC1); 1, 6, 12, 24 and 96 hours after transfusion (T1, T6, T12, T24 and T96 respectively); 8, 16 and 32 days after transfusion (T8d, T16d and T32d respectively). Before transfusion, blood samples were also withdrawn from the bags for the same analyzes. Statistical analyzes were performed in the statistical program GRAPHPAD PRISM 5.0, adopting a significance level of 5%. The bags of blood stored for 15 and 35 days showed more biochemical changes, blood gas, oxidative stress and lipid peroxidation than fresh blood bags. As for the biochemical analysis, after the transfusion was observed an increase of the total protein, albumin, glucose and creatine kinase in the 3 groups, and elevation of total bilirubin, direct bilirubin, and urea in G15 and G35. The changes observed in the blood gas analysis had no clinical significance, as they were within the reference values for the species. The goats that received stored blood showed disorder in their antioxidant system through alteration of the SOD activity. In the analysis of lipid peroxidation no difference between the groups for the concentration of malondialdehyde was found. Thus, it can be concluded that transfusion of whole fresh stored blood in goats did not compromise the blood gases, lipid peroxidation and liver and renal functions of the transfused animals. In addition, the method was proved to be efficient to restore, among other components, the total protein and albumin. The transfusion, as performed in this study, proved to be safe for used in the clinical practice of goats.(AU)

A transfusão de sangue é um procedimento terapêutico de grande importância para medicina veterinária, apesar disto, na literatura há poucos estudos sobre hemoterapia em caprinos. Este trabalho teve como objetivo avaliar as respostas bioquímicas, hemogasométricas, estresse oxidativo e peroxidação lipídica de caprinos submetidos à transfusão homóloga de sangue total fresco ou armazenado por 15 e 35 dias. Foram utilizados 18 caprinos adultos, machos, submetidos a uma única flebotomia para retirada de 30% do volume sanguíneo e transfundidos com 20mL/kg de sangue total armazenado em bolsas CPDA-1 de acordo com o grupo experimental, sendo: o G0 composto por animais que receberam sangue fresco; G15 e G35 animais que receberam sangue armazenado por 15 e 35 dias, respectivamente. Para a avaliação bioquímica, hemogasométrica, estresse oxidativo e peroxidação lipídica foram coletadas amostras de sangue nos seguintes momentos: antes da indução da anemia (TC0); 6 horas após a flebotomia e antes de transfusão (TC1); 1, 6, 12, 24 e 96 horas após a transfusão (T1, T6, T12, T24 e T96, respectivamente); 8, 16 e 32 dias após a transfusão (T8d, T16d e T32d, respectivamente). Antes da transfusão, amostras de sangue também foram retiradas das bolsas para realização das mesmas análises. As análises estatísticas foram realizadas no programa estatístico GRAPHPAD PRISM 5.0, adotando-se nível de significância de 5%. As bolsas de sangue armazenadas por 15 e 35 dias apresentaram mais alterações bioquímicas, hemogasométricas, estresse oxidativo e peroxidação lipídica do que as bolsas de sangue fresco. Quanto à análise bioquímica, após a transfusão observou-se aumento da proteína total, albumina, glicose e creatina quinase nos 3 grupos; e elevação da bilirrubina total, bilirrubina direta e ureia no G15 e G35. As alterações observadas na análise hemogasométrica não tiveram significado clínico, pois estavam dentro dos valores de referência para a espécie. Os animais que receberam sangue armazenado apresentaram desordem no seu sistema antioxidante demonstrada pela alteração da atividade da SOD. Na análise da peroxidação lipídica não houve diferença entre os grupos para a concentração de malondialdeído. Sendo assim, pode-se concluir que a transfusão de sangue homólogo total fresco ou armazenado em caprinos não comprometeu a gasometria, peroxidação lipídica e funções hepática e renal dos animais transfundidos. Além disso, mostrou-se eficiente em repor entre outros componentes, a proteína total e albumina. Desta forma, a transfusão, conforme realizada neste estudo, mostrou-se segura para ser utilizada na prática clínica desta espécie.(AU)

Potential prophylactic effect of recombinant bovine somatotropin (rbST) in sheep with experimentally induced hyperketonemia.

Hyperketonemia in sheep is associated with a disease known as pregnancy toxemia. The purposes of this study were to (1) induce hyperketonemia by infusion with D-beta-hydroxybutyrate (D-BHB) in eighteen healthy, non-pregnant, non-lactating ewes, (2) to evaluate the metabolic pathways in which recombinant bovine somatotropin (rbST) may be involved, and (3) to evaluate the possible benefits of preventive rbST therapy on the metabolic profiles of the sheep. The sheep were intravenously infused with a D-BHB solution over 2 h. Three days prior to the infusion, one group (n = 9) was injected with a single dose of 160 mg rbST, and the other group (n = 9) was injected with saline as a control. Blood samples were collected and metabolic profiles were analyzed every 20 min during the infusion and at 15, 30, 60, 120, 180, and 240 min post-infusion. Hyperketonemia was successfully achieved, demonstrated by serum BHB concentrations of 3.9 and 3.0 mmol/L in the rbST and control groups, respectively, without the presence of clinical signs. An increase in peripheral insulin resistance was observed after infusion but not during the infusion, in the rbST group. Higher glucose concentrations were observed in the rbST group post-infusion, suggesting a positive impact of D-BHB on gluconeogenesis. Our data suggest that prophylactic treatment with rbST may be useful in sheep to reduce the risk of PT.

Evidence of zoonotic leprosy in Pará, Brazilian Amazon, and risks associated with human contact or consumption of armadillos.

Mycobacterium leprae (M. leprae) is a human pathogen and the causative agent for leprosy, a chronic disease characterized by lesions of the skin and peripheral nerve damage. Zoonotic transmission of M. leprae to humans by nine-banded armadillos (Dasypus novemcinctus) has been shown to occur in the southern United States, mainly in Texas, Louisiana, and Florida. Nine-banded armadillos are also common in South America, and residents living in some areas in Brazil hunt and kill armadillos as a dietary source of protein. This study examines the extent of M. leprae infection in wild armadillos and whether these New World mammals may be a natural reservoir for leprosy transmission in Brazil, similar to the situation in the southern states of the U.S. The presence of the M. leprae-specific repetitive sequence RLEP was detected by PCR amplification in purified DNA extracted from armadillo spleen and liver tissue samples. A positive RLEP signal was confirmed in 62% of the armadillos (10/16), indicating high rates of infection with M. leprae. Immunohistochemistry of sections of infected armadillo spleens revealed mycobacterial DNA and cell wall constituents in situ detected by SYBR Gold and auramine/rhodamine staining techniques, respectively. The M. leprae-specific antigen, phenolic glycolipid I (PGL-I) was detected in spleen sections using a rabbit polyclonal antibody specific for PGL-I. Anti-PGL-I titers were assessed by ELISA in sera from 146 inhabitants of Belterra, a hyperendemic city located in western Pará state in Brazil. A positive anti-PGL-I titer is a known biomarker for M. leprae infection in both humans and armadillos. Individuals who consumed armadillo meat most frequently (more than once per month) showed a significantly higher anti-PGL-I titer than those who did not eat or ate less frequently than once per month. Armadillos infected with M. leprae represent a potential environmental reservoir. Consequently, people who hunt, kill, or process or eat armadillo meat are at a higher risk for infection with M. leprae from these animals.

Novel Anaplasma and Ehrlichia organisms infecting the wildlife of two regions of the Brazilian Amazon.

During 2009-2012, wild animals were sampled in the Amazon biome of Brazil. Animal tissues and blood were tested by polymerase chain reaction (PCR) assays targeting DNA of the bacterial family Anaplasmataceae (genera Anaplasma, Ehrlichia, Wolbachia) and the genus Borrelia. Overall, 181 wild animals comprising 36 different species (2 reptiles, 5 birds, and 29 mammals) were sampled. All birds and reptiles were negative by all PCR assays, as well as all mammals for the Borrelia PCR assay. Anaplasmataceae agents were searched by PCR assays targeting two different genes, the ribosomal 16S rRNA gene and the protein-coding dsb gene. Three dsb closely related haplotypes were generated from 3 white-lipped peccaries (Tayassu pecari). In a phylogenetic analysis inferred from dsb partial sequences, these haplotypes grouped with previously reported Ehrlichia haplotypes from jaguar (Panthera onca) and horse from Brazil, suggesting that they could all represent a single species, yet to be properly characterized. A unique dsb haplotype was generated from a sloth (Bradypus tridactylus), and could also represent a different Ehrlichia species. All these dsb haplotypes formed a clade sister to the Ehrlichia ruminantium clade. Three distinct 16S rRNA gene haplotypes were generated from a wild guinea pig (Cavia sp.), a woolly mouse opossum (Micoureus demerarae), and two from robust capuchin monkeys (Sapajus sp.). In a phylogenetic analysis inferred from 16S rRNA gene partial sequence, these haplotypes grouped within the Wolbachia clade, and are likely to represent Wolbachia organisms that were infecting invertebrate metazoarians (e.g., filarids) associated with the sampled mammals. Two deer (Mazama americana) samples yielded two distinct 16S rRNA gene sequences, one identical to several sequences of Anaplasma bovis, and an unique sequence that grouped in a clade with different Anaplasma species. Our results indicate that a variety of genetically distinct Anaplasmataceae organisms, including potentially new Ehrlichia species, circulate under natural conditions in the Amazonian wildlife.

Novel piroplasmid and Hepatozoon organisms infecting the wildlife of two regions of the Brazilian Amazon.

During 2009-2012, wild animals were sampled in two areas within the Amazon biome of Brazil, in the states of Mato Grosso and Pará. Animal tissues and blood were molecularly tested for the presence of Piroplasmida (genera Babesia, Theileria, Cytauxzoon) or Hepatozoon DNA. Overall, 181 wild animals comprising 36 different species (2 reptiles, 5 birds, and 29 mammals) were sampled. The following Piroplasmida agents were detected: Cytauxzoon felis in one ocelot (Leopardus pardalis), Theileria cervi in two red brocket deer (Mazama americana), Theileria spp. in three nine-banded-armadillos (Dasypus novemcinctus), one agouti (Dasyprocta sp.), and four lowland pacas (Cuniculus paca), Babesia spp. in one common opossum (Didelphis marsupialis) and one white-lipped peccary (Tayassu pecari). The following Hepatozoon agents were detected: Hepatozoon sp. (possibly Hepatozoon caimani) in three spectacled caimans (Caiman crocodilus), Hepatozoon felis in an ocelot (Leopardus pardalis), and Hepatozoon spp. in one scorpion mud turtle (Kinosternon scorpioides) and one lowland paca (Cuniculus paca). Phylogenetic analyses inferred by the 18S rRNA gene partial sequences supported these results, highlighting at least five novel Piroplasmida agents, and two novel Hepatozoon agents. This study screened the presence of tick-borne protozoa in a number of wildlife species from the Amazon for the first time. Our results indicate that a variety of genetically distinct Piroplasmida and Hepatozoon organisms circulate under natural conditions in the Amazonian wildlife.

Trypanosoma vivax in water buffalo of the Venezuelan Llanos: An unusual outbreak of wasting disease in an endemic area of typically asymptomatic infections.

Trypanosoma vivax has been associated with asymptomatic infections in African and South American buffalo. In this study, T. vivax was analyzed in water buffalo (Bubalus bubalis) from Venezuela in a molecular survey involving 293 blood samples collected from 2006 to 2015 across the Llanos region. Results demonstrated constant infections (average 23%) during the years analyzed. In general, animals were healthy carriers of T. vivax with low levels of parasitemia and were diagnosed exclusively by TviCATL-PCR. However, an outbreak of severe acute infections mostly in dairy animals was reported during a prolonged drought affecting 30.4% of a buffalo herd (115 animals examined). During the outbreak, animals exhibiting anemia and neurological disorders developed fatal infections, and 7% of the herd died within nine months before treatment against trypanosomosis. Microsatellite locus genotyping (MLG) of T. vivax samples before and during the outbreak revealed similar genotypes, but outbreak isolates exhibited the most divergent MLG. Venezuelan samples from symptomless and sick buffalo did not share the MLGs previously detected in asymptomatic Brazilian buffalo. Trypanosoma evansi was not detected in the herd examined during the outbreak. However, as expected Babesia sp. (62.6%) and Anaplasma sp. (55.6%) infections were highly prevalent in asymptomatic buffalo in the studied areas. This is the first South American outbreak of highly lethal acute T. vivax infections in water buffalo. Our results suggest that chronically infected and asymptomatic buffalo living in areas of enzootic equilibrium can develop symptomatic/lethal disease triggered by stressful scarcity of green forage and water during long droughts, inappropriate management of herds and likely concomitant anaplasmosis and babesiosis. Altogether, these factors weaken buffalo immune defenses, allowing T. vivax to proliferate and, consequently, allowing for progression to wasting disease.

Effects of sudden melon intake on ruminal parameters of non-adapted sheep / Efeitos da ingestão súbita de melão sobre os parâmetros ruminais de ovinos não adaptados

This study evaluated the effects of varying amounts of melon with high sugar content offered to sheep without prior melon experience and that were not adapted to consuming it. We used 12 eight-month-old, rumen-cannulated crossbred sheep weighing 25 kg each. The animals received a base diet of roughage, and then half were randomly selected to have 25% of their diet replaced with melon (G25%) and the other half had 75% of their diet replaced with melon (75%). Ruminal fluid was collected before administration of melon and at 0, 3, 6, 12, 18, and 24 h after the administration of the fruit. Sheep from the G25% group presented volatile fatty acid ruminal acidosis (sub-acute) between 3 and 6 h after consumption. This acidosis was characterized by a rumen pH slightly lower than 5.6, increased discrete L-lactic acid content, and increased redox potential (RP) and methylene blue redox (MBR) time of the ruminal fluid. The G75% group presented lactic ruminal acidosis at T6h, characterized by a rumen pH lower than 5.0, high lactate-L content, increased RP and MBR time, and increased ruminal fluid osmolarity. Therefore, offering large amounts of melon (75% of dry matter (DM)) is not recommended but 25% of DM of this fruit can be used safely.

O presente estudo avaliou o efeito da adição à dieta de duas diferentes quantidades de melão com alto teor de açucares oferecidos subitamente à ovinos não adaptados. Foram utilizados 12 ovinos mestiços da raça Santa Inês, com 8 meses de idade e pesando cerca der 25 kg providos de cânula ruminal. Os animais recebiam dieta à base de volumoso e foram aleatoriamente divididos em dois grupos iguais que receberam 25 ou 75% de adição de melão picado à dieta. Foram coletadas amostras de conteúdo ruminal nos momentos zero e após 3, 6, 12, 18, e 24 horas da administração do melão. Ovinos do grupo G25% apresentaram acidose por ácido graxo voláteis (subaguda) entre 3 e 6 horas, caracterizada por pH ruminal ligeiramente abaixo de 5,6, aumento discreto na concentração de ácido láctico L e aumento do potencial redox (PR) e tempo de redução do azul de metileno (TRAM) do fluido ruminal. O grupo G75% apresentou acidose láctica ruminal com pH menor que 5,0, alta concentração de Lactato-L e aumento do PR e TRAM e da osmolaridade ruminal. Em conclusão, o oferecimento de grandes quantidades de melão (75% da MS) não é recomendado, mas 25% da MS desta fruta pode ser utilizada na dieta de ovinos com segurança.

Anuran trypanosomes: phylogenetic evidence for new clades in Brazil.

Trypanosomes of anurans and fish are grouped into the Aquatic Clade which includes species isolated from fish, amphibians, turtles and platypus, usually transmitted by leeches and phlebotomine sand flies. Trypanosomes from Brazilian frogs are grouped within the Aquatic Clade with other anuran trypanosome species, where there seems to be coevolutionary patterns with vertebrate hosts and association to Brazilian biomes (Atlantic Forest, Pantanal and Amazonia Rainforest). We characterised the anuran trypanosomes from two different areas of the Cerrado biome and examined their phylogenetic relationships based on the SSU rRNA gene. A total of 112 anurans of six species was analysed and trypanosome prevalence evaluated through haemoculture was found to be 7% (8 positive frogs). However, only three isolates (2.7%) from two anuran species were recovered and cryopreserved. Analysis including SSU rDNA sequences from previous studies segregated the anuran trypanosomes into six groups, the previously reported An01 to An04, and An05 and An06 reported herein. Clade An05 comprises the isolates from Leptodactylus latrans (Steffen) and Pristimantis sp. captured in the Cerrado biome and Trypanosoma chattoni Mathis & Leger, 1911. The inclusion of new isolates in the phylogenetic analyses provided evidence for a new group (An06) of parasites from phlebotomine hosts. Our results indicate that the diversity of trypanosome species is underestimated since studies conducted in Brazil and other regions of the world are still few.

Ticks and rickettsial infection in the wildlife of two regions of the Brazilian Amazon.

During 2009-2012, wild animals and their ticks were sampled in two areas within the Amazon biome of Brazil, in the states of Mato Grosso and Pará. Animal tissues, blood, and ticks were molecularly tested for Rickettsia and Coxiella DNA. A total of 182 wild animals were sampled, comprising 28 mammalian, five avian, and three reptilian species. Animal tissues or blood were all negative for Rickettsia or Coxiella DNA. A total of 454 ticks (22 larvae, 226 nymphs, 127 males, 79 females) were collected from 52 (28.6%) animals, and identified into 15 species: Amblyomma cajennense, A. naponense, A. humerale, A. nodosum, A. goeldii, A. oblongoguttatum, A. longirostre, A. calcaratum, A. coelebs, A. pacae, A. geayii, A. rotundatum, A. auricularium, A. ovale, and Haemaphysalis juxtakochi. While no Coxiella DNA was identified in ticks, six Rickettsia species were detected in the ticks. "Candidatus Rickettsia amblyommii" was the most common agent, detected in four tick species, A. cajennense, A. auricularium, A. longirostre, and A. humerale. The second most common agent, R. bellii, was detected in A. humerale and A. naponense. Rickettsia rhipicephali was detected in H. juxtakochi, and R. felis in A. humerale. Two possible new Rickettsia species were detected in A. naponense ticks, namely, a novel spotted fever group agent close-related to R. africae in Pará, and a novel Canadensis group agent in Mato Grosso. Results of the present study expand our knowledge on the tick fauna, and on the yet infantile knowledge of tick-borne rickettsiae in the Amazon biome.

Comparison of rumen fluid pH by continuous telemetry system and bench pH meter in sheep with different ranges of ruminal pH.

We aimed to compare the measurements of sheep ruminal pH using a continuous telemetry system or a bench pH meter using sheep with different degrees of ruminal pH. Ruminal lactic acidosis was induced in nine adult crossbred Santa Ines sheep by the administration of 15 g of sucrose per kg/BW. Samples of rumen fluid were collected at the baseline, before the induction of acidosis (T 0) and at six, 12, 18, 24, 48, and 72 hours after the induction for pH measurement using a bench pH meter. During this 72-hour period, all animals had electrodes for the continuous measurement of pH. The results were compared using the Bland-Altman analysis of agreement, Pearson coefficients of correlation and determination, and paired analysis of variance with Student's t-test. The measurement methods presented a strong correlation (r = 0.94, P < 0.05) but the rumen pH that was measured continuously using a telemetry system resulted in lower values than the bench pH meter (overall mean of 5.38 and 5.48, resp., P = 0.0001). The telemetry system was able to detect smaller changes in rumen fluid pH and was more accurate in diagnosing both subacute ruminal lactic acidosis and acute ruminal lactic acidosis in sheep.

Phylogenetic relationships of Leishmania species based on trypanosomatid barcode (SSU rDNA) and gGAPDH genes: Taxonomic revision of Leishmania (L.) infantum chagasi in South America.

Phylogenetic studies on trypanosomatid barcode using V7V8 SSU rRNA and gGAPDH gene sequences have provided support for redefining some trypanosomatid species and positioning new isolates. The genus Leishmania is a slow evolving monophyletic group and including important human pathogens. The phylogenetic relationships of this genus have been determined by the natural history of its vertebrate hosts, vector specificity, clinical manifestations, geographical distribution and molecular approaches using different markers. Thus, in an attempt to better understand the phylogenetic relationships of Leishmania species, we performed phylogenetic analysis on trypanosomatid barcode using V7V8 SSU rRNA and gGAPDH gene sequences among a large number of Leishmania species and also several Brazilian visceral Leishmania infantum chagasi isolates obtained from dogs and humans. Our phylogenetic analysis strongly suggested that Leishmania hertigi and Leishmania equatoriensis should be taxonomically revised so as to include them in the genus Endotrypanum; and supported ancient divergence of Leishmania enriettii. This, together with recent data in the literature, throws light on the discussion about the evolutionary southern supercontinent hypothesis for the origin of Leishmania ssp. and validates L. infantum chagasi from Brazil, thus clearly differentiating it from L. infantum, for the first time.

Isolation and phylogenetic relationships of bat trypanosomes from different biomes in Mato Grosso, Brazil.

In the order Chiroptera, more than 30 trypanosome species belonging to the subgenera Herpetosoma, Schizotrypanum, Megatrypanum, and Trypanozoon have been described. The species Trypanosoma cruzi , Trypanosoma cruzi marinkellei, and Trypanosoma dionisii are the most common in bats and belong to the Schizotrypanum subgenus. Bats from 2 different biomes, Pantanal and Amazonia/Cerrado in the state of Mato Grosso, Brazil, were evaluated according to the presence of trypanosome parasites by means of hemoculture and PCR in primary samples (blood samples). A total of 211 bats from 20 different species were caught and the trypanosome prevalence, evaluated through hemoculture, was 9.0% (19), 15.5% (13), and 4.8% (6) in the municipalities of Confresa (Amazonia/Cerrado biome) and Poconé (Pantanal biome). Among the 123 primary samples obtained from the bats, only 3 (2.4%) were positive. Phylogenetic analysis using trypanosomatid barcoding (V7V8 region of SSU rDNA) identified all the isolates and primary samples as T. c. marinkellei. The sequences of the isolates were segregated according to the bat host genus or species and suggest that co-evolutionary patterns exist between hosts and parasites. Further studies in different Brazilian regions and biomes need to be conducted in order to gain real understanding of the diversity of trypanosomes in bats.

Evaluation of hematologic, blood gas, and select biochemical variables in ovine whole blood stored in CPDA-1 bags.

BACKGROUND: The economic consequences from mortality of sheep after blood loss can be considerable. To date there are no reports addressing hematologic, blood gas, and biochemical changes in ovine blood stored in CPDA-1 bags. OBJECTIVES: The aim of this study was to investigate hematologic, blood gas, and biochemical alterations resulting from storage of ovine blood in CPDA-1 bags to establish transfusion protocols in sheep. METHODS: From each of 7 healthy 8-month-old sheep 450 mL of blood were collected into CPDA-1 bags and stored for 35 days in at 3-6°C. Samples were taken from the bags at days 0, 7, 14, 21, and 35. Whole blood was used to assess PCV, MCV, RBC count, pH, pO2 , pCO2 , and concentrations of bicarbonate, sodium, and lactate. Plasma was used to measure potassium, hemoglobin, and glucose concentrations. RESULTS: The PCV remained stable throughout the storage period, while plasma hemoglobin and MCV began to increase on days 7 and 21, respectively. The RBC count began to decrease on day 21. Blood pH decreased and pCO2 increased steadily throughout the storage period. Potassium concentration increased from 3.8 to 18.3 mmol/L on day 7 and remained high thereafter. In contrast, sodium concentration began to decrease on day 7. CONCLUSIONS: The results show that ovine blood undergoes hematologic, blood gas, and biochemical changes during storage. Further studies are required to establish RBC viability in CPDA-1 bags after a storage period of 35 days.

First Report of Trypanosoma sp. in Spectacled Caiman (Caiman crocodilus): Morphological and Phylogenetic Relationships.

In Crocodylidae family three trypanosomes species were described, T. grayi in African crocodilian and T. cecili and Trypanosoma sp. in Caimans species from Brazil. T. grayi was transmitted by tsetse flies and the vector of Brazilian caimans trypanosomes is unknown. We characterized first Brazilian trypanosome isolated in spectacled caiman (Caiman crocodilus) from Mato Grosso State in Brazil. Morphological findings in epimastigotes forms from axenic culture showed high similarity with Trypanosoma sp. described in Caiman yacare from Brazilian Pantanal. Phylogenetic studies performed with SSU rDNA and gGAPDH (glyceraldehydes-3-phosphato dehydrogenase glycosomal) clustering in T. grayi Clade and together to genotype Cay 01 from Trypanosoma unnamed species isolated in C. yacare. This is the first isolate of Trypanosoma sp. from C. crocodilus and the phylogenetic position with isolates in C. yacare from Pantanal region and demonstrates the low host specificity of cayman trypanosomes in Brazil.

Molecular confirmation of ovine herpesvirus 2-induced malignant catarrhal fever lesions in cattle from Rio Grande do Norte, Brazil / Diagnóstico molecular de herpesvírus ovino tipo 2 em surto de febre catarral malígna em bovinos do Rio Grande do Norte

Molecular findings that confirmed the participation of ovine herpesvirus 2 (OVH-2) in the lesions that were consistent with those observed in malignant catarrhal fever of cattle are described. Three mixed-breed cattle from Rio Grande do Norte state demonstrated clinical manifestations that included mucopurulent nasal discharge, corneal opacity and motor incoordination. Routine necropsy examination demonstrated ulcerations and hemorrhage of the oral cavity, corneal opacity, and lymph node enlargement. Significant histopathological findings included widespread necrotizing vasculitis, non-suppurative meningoencephalitis, lymphocytic interstitial nephritis and hepatitis, and thrombosis. PCR assay performed on DNA extracted from kidney and mesenteric lymph node of one animal amplified a product of 423 base pairs corresponding to a target sequence within the ovine herpesvirus 2 (OVH-2) tegument protein gene. Direct sequencing of the PCR products, from extracted DNA of the kidney and mesenteric lymph node of one cow, amplified the partial nucleotide sequences (423 base pairs) of OVH-2 tegument protein gene. Blast analysis confirmed that these sequences have 98-100% identity with similar OVH-2 sequences deposited in GenBank. Phylogenetic analyses, based on the deduced amino acid sequences, demonstrated that the strain of OVH-2 circulating in ruminants from the Brazilian states of Rio Grande do Norte and Minas Gerais are similar to that identified in other geographical locations. These findings confirmed the active participation of OVH-2 in the classical manifestations of sheep associated malignant catarrhal fever.

Os achados moleculares confirmaram a participação do herpesvírus ovino tipo 2 (OVH-2) nas lesões observadas em um surto de febre catarral malígna em bovinos. Três bovinos oriundos de propriedade rural de Mossoró, Rio Grande do Norte apresentaram manifestações clínicas, que incluíram secreção nasal mucopurulenta, opacidade da córnea e incoordenação motora. A necropsia revelou ulcerações e hemorragias da cavidade oral, opacidade da córnea e linfonodomegalia. Os achados histopatológicos significativos incluíam vasculite necrosante generalizada, meningoencefalite não supurativa, nefrite intersticial linfocítica, hepatite linfocítica e trombose. A PCR, realizada a partir de DNA extraído do rim e do linfonodo mesentérico de um dos animais, amplificou um produto com 423 pares de base do gene da proteína do tegumento do herpesvírus ovino 2 (OVH-2). O sequenciamento direto dos produtos da PCR e a análise pelo Blast demonstraram que o produto amplificado apresentava 98-100% de identidade com sequências do OVH-2 depositadas no GenBank. As análises filogenéticas, baseadas nas sequências deduzidas de aminoácidos demonstraram que a cepa de OVH-2 circulando em ruminantes nos estados de Rio Grande do Norte e Minas Gerais são semelhantes àquelas identificadas em outras regiões geográficas. Esses achados confirmam a participação ativa de OVH-2 nas manifestações clássicas de febre catarral maligna em ovinos.

Prevalence of anti- Neospora caninum and anti- Toxoplasma gondii antibodies in dogs from two different indigenous communities in the Brazilian Amazon Region.

Prevalence of Toxoplasma gondii and Neospora caninum antibodies in sera of 325 dogs in 11 villages inhabited by the Tapirapé and Karajá ethnic groups in the south of the Brazilian Amazon was determined by the use of an indirect fluorescence antibody test. Antibodies (cutoff 1:16) to T. gondii were found in 169 (52%) and to N. caninum (cut-off 1:50) in 32 (9.8%) of 325 dogs. Seropositivity for both parasitic infections was widely prevalent in dogs from these villages and was higher in older dogs, indicating post-natal transmission.

Occurrence of Toxoplasma gondii antibodies in Dasyprocta aguti from Brazil: comparison of diagnostic techniques.

In this study, the occurrence of antibodies to Toxoplasma gondii in Brazilian agouti (Dasyprocta aguti) was compared by modified agglutination test (MAT) and indirect fluorescent antibody test (IFAT) using anti-capybara conjugate. Sera from 109 animals were tested using MAT (1:25 cut-off) and IFAT (1:16 cut-off); 19% were positive by MAT, and 18% were positive by IFAT. Overall, the 17 IFAT-positive samples were also positive for MAT. The four positive MAT samples with a titer < or = 200 were IFAT negative. All negative samples obtained by MAT matched with the results of the IFAT. Comparing both tests, and considering MAT as the gold standard, the sensitivity of IFAT was 81%, the specificity was 100%, the accuracy was 97%, the positive predictive value (PPV) was 100%, and the negative predictive value 96%. The kappa value agreement was 87.3% (75.1-99.6%). The anti-capybara conjugate can be successfully used to perform IFAT in Brazilian agouti with maximum specificity and PPV.