Occurrence and risk factors associated with gastrointestinal parasitism in horses reared in different systems.

To facilitate exploration of the immense range of gastrointestinal parasites in horses and the impact of parasitism on equine health, the present study aimed to evaluate the occurrence and risk factors associated with these infections in horses reared in different management systems in the state of Santa Catarina, Brazil. Samples were collected from 208 horses: 91 from extensive, 64 from semi-extensive, and 53 from intensive rearing systems. The identified helminths included those of the Strongylida order (80.29%), Parascaris equorum (3.36%), Oxyuris equi (4.33%), and Anoplocephala spp. (1.92%). By analyzing the coproculture results to differentiate among Strongylida order parasites, species such as Strongylus vulgaris, S. edentatus, S. equinus, Triodontophorus spp., and Trichostrongylus axei, in addition to members of the Cyathostominae subfamily, which include Gyalocephalus capitatus and Poteriostomum spp., were identified. The only positive sample of protozoa was that of Cryptosporidium spp. (13%). Regarding the rearing system, animals in the extensive system had a higher proportion of infected horses and a higher chance of infection than those in the other systems. For the variable co-grazing with cattle, only cyathostomins showed a significant difference with relatively low infection risk in co-grazing. Overall, the present study demonstrated a high occurrence of equine gastrointestinal parasites, especially those of the Strongylida order, with emphasis on small strongylids. In addition, analyzing factors associated with infection showed that management variables are important for controlling parasitism in horses.

Interaction between live vaccines for coccidiosis and phytogenic compounds in the diet of broilers.

This study aimed to determine whether live vaccines alone or associated with phytogenic compounds supplied via feed could improve performance, intestinal health, serum biochemistry, and coccidiostatic action in broilers up to 42 days of age, challenged with Eimeria spp., compared with a traditional anticoccidial program. We used 800 birds divided into five treatments of eight repetitions each (n = 20): NC - Negative control (absence of additives in the feed and without coccidian challenge); PC - Positive control (absence of additives and with coccidian challenge at 21 days); AAP - Anticoccidial additives program, including salinomycin and nicarbazine, with coccidian challenge at 21 days of life; VAC - Vaccination on day 1 of life (Hipracox HIPRA®) against coccidiosis and subsequent coccidian challenge at 21 days; VAC + BPC - Vaccination (day 1 of life), the addition of 200 ppm of blend phytogenic compound (BPC) based on carvacrol and cinnamaldehyde to the diet, with coccidian challenge at 21 days of life. The birds in the AAP group more significant weight gain and greater body weight at 21 days than the NC and 42 days of age than the NC and VAC + BPC groups. The excretion of Eimeria oocysts in the excreta of birds was higher in PC than in NC, and in the litter, PC had higher counts than NC, AAP, and VAC + BPC. A higher mean score of intestinal lesions for Eimeria acervulina was observed in VAC and VAC + BPC birds, while for Eimeria maxima, the highest score was found in PC. Higher serum levels of total protein due to the increase in globulins were observed in birds of AAP and VAC than NC. Our findings suggest that vaccination for coccidiosis shortly after the birth of chicks allows performance similar to the conventional coccidiosis control program via diet.

Effect of a phytogenic feed additives mixture on milk physico-chemical properties and biochemical parameters of Holstein cows / Efeito de uma mistura de aditivos fitogênicos alimentares nas propriedades físico-químicas do leite e parâmetros bioquímicos de vacas leiteiras da raça holandês

ABSTRACT: This study aimed to analyze the effect of phytogenic additives based on cardol, cardanol, and ricinoleic acid on the physico-chemical qualities and biochemical parameters of Holstein cow milk. Nineteen animals were divided into the control (GI) and treatment (GII) groups. Prior to the beginning of the experiment, sodium monensin was removed from the feed provided to the animals. This study consisted of two phases. In the first, animals from GI received the standard diet, without additives, while animals from GII received the standard diet supplemented with 10 g of phytogenic additives for 60 days. At the end of the first stage, the animals remained 30 days without receiving additives and in the second phase the groups were inverted, and the GII again received the phytogenic additive during the subsequent 60 days. Physical-chemical analyzes of milk were performed at moments 0, 15, 30, 45 and 60 days of the experiment and the blood parameters at times 0 and 60 days of the experiment were evaluated. There was no significant difference for the treatment x time interaction in any of the variables, but there was a trend between groups for the CCS parameter and there was a statistical difference for the protein, as well as for the AST. In this study it is observed that the addition of phytogenic additive modulating rumen fermentation based on cardol, cardanol and ricinoleic acid does not compromise, in general, the quality of milk, with positive results for some specific parameters such as protein content and aminotransferase.

RESUMO: Esse estudo teve por objetivo avaliar o efeito de um aditivo fitogênico a base de cardol, cardanol e ácido ricinoléico na qualidade físico-química do leite e perfil bioquímico de vacas da raça Holandês. Dezenove animais foram divididos em dois grupos: Grupo Controle (GI) e Grupo Tratado (GII). Previamente ao início do experimento, retirou-se a monensina sódica da ração fornecida aos animais. Este estudo consistiu de duas fases, sendo que na primeira fase, os animais do GI receberam a dieta padrão, sem aditivos, enquanto os animais do grupo GII receberam a ração padrão suplementada com 10 g dos aditivos fitogênicos durante 60 dias. Ao final da primeira etapa, os animais permaneceram 30 dias sem receber aditivos e na segunda fase os grupos foram invertidos, sendo que o GII novamente recebeu o aditivo fitogênico durante os 60 dias subsequentes. Foram realizadas análises físico-químicas do leite nos momentos 0, 15, 30, 45 e 60 dias do experimento e avaliados os parâmetros sanguíneos nos momentos 0 e 60 dias do experimento. Não se observou diferença significativa para a interação tratamento x tempo em nenhuma das variáveis, mas tendência entre grupos para o parâmetro de CCS, e houve diferença estatística para a proteína, assim como para a AST. Neste estudo observa-se que a adição de aditivo fitogênico modulador de fermentação ruminal a base de cardol, cardanol e ácido ricinoléico não compromete, de forma geral, a qualidade do leite, tendo resultados positivos para alguns parâmetros específicos como teor de proteína e aspartato aminotransferase.

Factors associated with the efficiency of acaricides on different populations of Rhipicephalus microplus / Fatores associados à eficiência de acaricidas sobre diferentes populações de Rhipicephalus microplus

Tick infestation causes major problems in cattle. Tick parasitism accounts for significant economic losses in many beef and dairy herds in the vast majority of the states in the Brazilian territory including the State of Santa Catarina in the southern region of the country. Tick resistance to several active principles occurs due to a number of factors including the indiscriminate, injudicious and inadequate use of tick insecticides. Considering the great importance of fighting off tick infestations in cattle, we evaluated the efficiency of 8 different topical active principles against the ixodid tick R. microplus and identified the main factors that contribute to the development of ticks that are resistant to acaricides in the farms and bovine herds studied. For such purpose, R. microplus telegionae were collected in 39 farms locates in different municipalities of the West of Santa Catarina, south Brazil. At the time of sample collection, information about the management, the history of the acar used in the herd, the number of annual applications of these pesticides, the frequency of acaricide rotation, and the frequency of technical monitoring for strategic tick control were retrieved. We collected this data in order determine contributing factors to the development resistant R. microplus populations. For the sensitivity profile of these ticks to a number of different acaricides tested, 10 engorged females for each principle and a control were used. The results of our study show that most associations between pyrethroids and organophosphates had an efficiency between 96.6% and 100% in the control of R. microplus except for the combination of Alfacipermetrine 15%, Ethion 16%, and Chlorpyrifos 8.5% that had 93.4% of efficiency. Among the chemicals not associated with any other type of compound, Amitraz 12.5% ​​was effective in 98.3% of the cases. In contrast, Cypermethrin 15% had an efficiency in only 25.8% of the farms/herds analyzed. Based on the results of the present study, we may infer that the methods of control and management used by producers may be related to the low indexes of resistance to topical acaricides in the different populations of ticks in the study area.

A infestação por carrapatos em bovinos gera grandes problemas, sendo esse parasito responsável por elevados prejuízos econômicos em diversos rebanhos de produção de corte e também de leite na maioria das regiões brasileiras, inclusive em Santa Catarina. A resistência dos carrapatos frente aos diversos princípios ativos pode ser decorrente de fatores como o uso indiscriminado de carrapaticidas, bem como a inadequada forma de aplicação dos mesmos. Tendo em vista a grande importância em combater essas infestações, buscou-se avaliar a eficiência de oito diferentes princípios ativos de contato contra o carrapato Rhipicephalus microplus, bem como identificar os principais fatores que possam contribuir com a seleção de carrapatos resistentes nas propriedades avaliadas. Para isso, teleóginas de R. microplus foram coletadas em 39 propriedades de diferentes municípios do Oeste de Santa Catarina. No momento da coleta também foram obtidas informações referentes ao sistema de manejo dos animais, histórico dos carrapaticidas utilizados, número de aplicações anuais, frequência de rodízios de acaricidas, e frequência de acompanhamento técnico para controle estratégico, com a finalidade de evidenciar os diferentes fatores que possam estar contribuindo para o surgimento de eventual resistência das populações do R. microplus. A avaliação de suscetibilidade foi realizada através da técnica de biocarrapaticidograma, utilizando 10 teleóginas ingurgitadas para cada princípio e mais o controle. Dentre os principais resultados obtidos, a maioria das associações entre piretróides e organofosforados demonstraram ter eficiência entre 96,6% e 100%, com exceção da associação de Alfacipermetrina 15%, Ethion 16% e Clorpirifós 8,5% com 93,4% de eficiência. Dentre as bases químicas sem nenhum tipo de associação o Amitraz 12,5% demonstrou ser eficaz em 98,3%, em contrapartida a Cipermetrina 15% teve eficiência em apenas 25,8% das propriedades analisadas. A partir dos resultados obtidos, pode se afirmar que as formas de controle e manejo adotados pelos produtores podem estar relacionadas com os baixos índices de resistência aos carrapaticidas de contato, nas diferentes populações de carrapato na região do estudo.

Prevalência de anticorpos e fatores de risco associados à infecção por Toxoplasma gondii em galinhas domésticas da zona rural de Santa Maria, Rio Grande do Sul / Prevalence of antibodies and risk factors associated with infection by Toxoplasma gondii in free range chickens of rural area of Santa Maria, Rio Grande do Sul, Brazil

RESUMO: Toxoplasma gondii é um protozoário apicomplexa que infecta animais de sangue quente, podendo ser considerado um dos principais parasitas capazes de infectar os seres humanos. Galinhas domésticas podem ser facilmente infectadas por protozoários, uma vez que estas podem ingerir oocistos encontrados no solo, sendo consideradas boas indicadoras de contaminação ambiental por T. gondii. O objetivo deste estudo foi determinar a presença de anticorpos anti-T. gondii em galinhas domésticas criadas extensivamente e avaliar os fatores de risco associados ao protozoário, na zona rural de Santa Maria, RS, Brasil. No período de março de 2013 a fevereiro 2014 foram coletadas 597 amostras de sangue de galinhas domésticas em 74 propriedades, oriundas de nove estratos que representam cada distrito da zona rural. Para avaliar os fatores de risco, nessas propriedades foi aplicado um questionário epidemiológico aos moradores. As amostras de soro foram testadas por imunofluorescência indireta, e 49,2% (294/597) foram positivas para anticorpos anti-T. gondii, com títulos variando de 16 a 4096. Das 74 propriedades analisadas, em 63 (85,1%) houve relatos que os gatos têm acesso ao deposito de alimentos, com associação significativa quando associado à presença de galinhas positivas (p=0,04) e o OR de 4,07. A variável "abate de animais" (aves e bovinos), em 51 (68,9%) das propriedades foi relatado o abate de bovinos e aves na propriedade, com valor de p significativo (p=0,05). A maioria das propriedades 59 (79,7%) foi relatada a presença de gatos domésticos, o que poderia estar associada com a alta soroprevalência encontrada em galinhas e a taxa de contaminação ambiental. A elevada prevalência de anticorpos encontrada neste estudo, além da alta frequência de propriedades com casos positivos, sugere uma grande contaminação ambiental nos distritos pesquisados, sendo assim um risco potencial para a saúde humana e animal.

ABSTRACT: Toxoplasma gondii is an apicomplex protozoan that infects warm-blooded animals and can be considered a major parasite capable of infecting humans. Domestic chickens can be easily infected by protozoa, since they can ingest oocysts found in the soil and are considered good indicators of environmental contamination by T. gondii. The aim of this study was to determine the presence of anti-T. gondii antibodies in free range chickens and to evaluate the risks factors associated with the protozoan in rural area of Santa Maria, RS, Brazil. From March 2013 to February 2014, 597 blood samples from domestic chickens were collected from 74 farms, from nine layers representing each district in the rural area. To evaluate the risk factors, in these farms an epidemiological questionnaire was applied to the residents. Serum samples were tested by indirect immunofluorescence and 49.2% (294/597) were positive for anti-T.gondii antibodies, with titres varying from 16 to 4096. Of the 74 analyzed farms, 63 (85.1%) reported that cats had access to food deposits, with a significant association when positive chickens were present (p = 0.04) and the OR of 4.07. The variable "slaughter of animals" (poultry and cattle) in 51 (68.9%) of the farms was reported the slaughter of cattle and birds in the farm, with significant p value (p = 0.05). Most farms 59 (79.7%) reported the presence of domestic cats, which could be associated with the high seroprevalence found in chickens and the rate of environmental contamination. The high prevalence of antibodies found in this study, in addition to the high frequency of farms with positive cases, suggests a great environmental contamination in the studied districts, thus being a potential risk to human and animal health.

Virulência e multiplicação de isolados de Toxoplasma gondii da região central do Rio Grande do Sul / Virulence and multiplication of Toxoplasma gondii isolates from the central region of Rio Grande do Sul, Brazil

O protozoário Toxoplasma gondii possui a capacidade de infectar diversas espécies animais, geralmente causando distúrbios reprodutivos. Quatro diferentes isolados de T. gondii - Pains #1 (P1), Pains #2 (P2), Santa Flora #1 (SF1) e Santa Flora #306 (SF306) - foram avaliados, após prévia genotipagem. A capacidade de multiplicação e virulência destes isolados foi analisada in vitro (células Vero) e in vivo (camundongos), sendo realizadas 3 passagens para cada isolado em cada modo avaliado, sendo sempre inoculada a dose de 1x104 taquizoítos em todas as passagens. Os camundongos eram observados diariamente, quanto à presença de sinais clínicos e ocorrência de mortalidade após inoculação dos taquizoítos. Os isolados SF1 e SF306, foram os que apresentaram maior multiplicação média do número total de taquizoítos em cada uma das 3 diferentes passagens realizadas para cada um dos isolados tanto in vitro quanto in vivo. Os primeiros sinais clínicos observados nos camundongos ocorreram entre os dias 5 a 11, após inoculação, com mortalidade acontecendo entre os dias 6 a 15, após inoculação. Assim, a multiplicação parasitária in vitro é semelhante à multiplicação in vivo destes isolados de T. gondii; diferentes isolados com o mesmo genótipo apresentam comportamento de virulência semelhante, caracterizando o isolado SF1 como mais virulento para camundongos.(AU)

The protozoan Toxoplasma gondii has the ability to infect several animal species, usually causing reproductive disorders. Four different isolates of T. gondii - Pains #1 (P1), Pains #2 (P2), Santa Flora #1 (SF1) and Santa Flora #306 (SF306) were evaluated with prior genotyping. Their virulence and multiplication capacity were analyzed in vitro (Vero cells) and in vivo (mice). For each isolate, three passages were performed with dose inoculation 1x104 tachyzoites at each passage. The mice were daily observed to verify of clinical signs and occurrence of mortality after tachyzoites inoculation. The SF1 isolate and SF306 isolate, presented the highest multiplication of the total tachyzoites number in each of the 3 different passages performed in vitro and in vivo. The initial clinical signs in mice were observed between 5 and 11 days, and occurring mortality between 6 and 15 days, after inoculation. Thus, the parasitic multiplication of theses isolates is similar in vitro and in vivo; different isolates within the same genotype have a similar virulence and the SF1 isolate is the most virulent for mice.(AU)

Sarcocystis species identification in cattle hearts destined to human consumption in southern Brazil.

Sarcocystis spp. are ubiquitous protozoan parasites that can form cysts in striated muscle and CNS of cattle. Cattle hearts are commonly infected by microscopic sarcocysts. Humans can get infected by consuming cattle meat containing the zoonotic parasites Sarcocystis hominis and Sarcocystis heydorni. However, bovine myocardium is generally infected by Sarcocystis cruzi. The aims of this study were to investigate the occurrence of sarcocysts and the identity of Sarcocystis species present in cattle hearts destined to human consumption in the Central region of the Rio Grande do Sul state, southern Brazil. A total of 314 cattle myocardium samples collected from a local abattoir were microscopically examined for the presence of sarcocysts. The sarcocysts isolated from 134 of these samples (ten sarcocysts per sample) were subjected to DNA extraction and PCR amplification. The PCR-amplified DNA fragments were digested with the restriction enzymes BclI and RsaI (PCR-RFLP) for differentiation among S. cruzi, S. hirsuta, and S. hominis. Sarcocystis species identification was confirmed using DNA sequencing of the cox1 mitochondrial DNA. Sarcocysts were detected in all the bovine myocardium samples. PCR-RFLP analysis resulted in successful amplification of 78 of the 134 samples tested. Only the S. cruzi DNA restriction pattern was identified from all of the 78 amplified samples. DNA sequencing also confirmed the presence of S. cruzi DNA. In conclusion, all myocardium samples evaluated were infected with microscopic sarcocysts. S. cruzi was the only species detected infecting the cattle hearts.

Diphenyl diselenide modulates nucleotidases, reducing inflammatory responses in the liver of Toxoplasma gondii-infected mice.

The aim of this study was to verify the effect of diphenyl diselenide (PhSe)2 on hepatic nucleotidases and on the concentration of purines in mice infected by Toxoplasma gondii. The animals were divided into four groups: Group A (uninfected), Group B (uninfected and treated with (PhSe)2), Group C (infected), and Group D (infected and treated with (PhSe)2). The inoculation (groups C and D) was performed with 50 cysts of T. gondii (ME-49 strain). Mice from groups B and D were treated with 5 µmol kg-1 of (PhSe)2. Liver tissue from infected mice showed less severe inflammation, elevated ATP/ADO ratio, elevated NTPDase, 5'nucleotidase, and ADA activities compared to the uninfected group (Group A; P < 0.05). However, infected and treated mice showed decreased ATP levels and elevated ADO levels, as well as higher NTPDase and 5'nucleotidase activities and decreased ADA activity in the hepatic tissue compared to the infected group (P < 0.05). Moreover, the (PhSe)2 treatment of infected mice reduced the hepatic inflammation and showed an immunomodulatory effect on ectonucleotidases of hepatic lymphocytes, which it returned to basal levels. Therefore, chronic infection by T. gondii induces hepatic inflammation in mice, and it is possible that purine levels and nucleotidase activities in hepatic tissue are related to the pathogenesis of the infection in this tissue. The treatment with (PhSe)2 was able to reverse the hepatic inflammation in mice chronically infected, possibly due to the modulation of purinergic enzymes that produce an anti-inflammatory profile through the purinergic system in the liver tissue.

Hepatic xanthine oxidase activity and purine nucleosides levels as physiological mediators to analyze a subcutaneous treatment with (PhSe)2 in mice infected by Toxoplasma gondii.

The aim of this study was to evaluate the levels of purine nucleosides and xanthine oxidase (XO) activity in the liver of mice chronically infected by Toxoplasma gondii and treated with diphenyl diselenide (PhSe)2. For this experiment, forty Swiss mice were used. Twenty animals were orally infected by approximately 50 bradizoites of a cystogenic ME-49 strain of T. gondii, and the same number of uninfected mice was used as a control group. Ten infected and ten uninfected mice were subcutaneously treated twice (days 1 and 20 post-infection (PI)) with 5 µmol kg-1 of (PhSe)2. On day 30 PI, liver samples were collected to measure the levels of hypoxanthine (HYPO), xanthine (XAN), uric acid (UA), and XO activity. Infected animals showed increased (P < 0.05) levels of hepatic XAN and UA, as well as XO activity compared to uninfected animals. The use of (PhSe)2 in healthy mice increased the levels of all nucleosides, but decreased XO activity compared to healthy untreated animals. The group of infected and treated animals showed increased XAN and UA levels, and XO activity compared to the healthy control group, however infected and treated mice showed a decrease in the XO activity compared to the infected untreated group. We conclude that chronic infection caused by T. gondii can induce hepatic changes, such as increased UA levels and XO activity, that can increase the pro-oxidative profile. The (PhSe)2 treatment of healthy animals altered the levels of nucleosides, possibly due to low XO activity that decreased nucleoside degradation. Finally, (PhSe)2 treatment decreased XO activity in the infected group and increased nucleoside levels; however it was unable to reduce the UA levels found during the infection.

Virulence factors and antimicrobial susceptibility profile of extraintestinal Escherichia coli isolated from an avian colisepticemia outbreak.

In this study an avian colisepticemia outbreak was investigated. Two isolates from a chicken with colisepticemia were characterized for antimicrobial susceptibility and virulence factors profile. For this purpose 7 antimicrobial and 29 genes (fimH, hrlA/hek, iha, papC, sfa/focCD, tsh, mat, tia, gimB, ibeA, chuA, fyuA, ireA, iroN, irp2, iucD, sitD. chr., sitD. ep., iss, neuC, ompA, traT, astA, hlyA, sat, vat, pic, malX, cvi/cva) were tested. The outbreak happened in a hick chicken breeding located in the northwestern region of Rio Grande do Sul state in South of Brazil and caused 28.3% (102 deads of a total of 360 chickens) of mortality rate. Escherichia coli isolates obtained from the avian spleen and liver belong to the same phylogenetic group A and present resistance to all antimicrobials tested (ampicillin, tetracycline, gentamicin, neomycin, sulfa + trimethoprim, enrofloxacin, and norfloxacin). Both isolates harbor virulence factors related to adhesion (fimH, papC, mat), invasion (tia), iron acquisition system (iroN) and serum resistance (iss, ompA, traT), showing that these groups are important for Avian Pathogenic E. coli (APEC). However, they present different virulence profiles for some genes, whereas liver-isolate carries more hrlA/hek (adhesin), gimB (invasin), sitD ep. (iron acquisition system), sat (toxin) and hylA (toxin) genes, the spleen-isolate harbors fyuA (iron acquisition system) gene. Here, we highlight a coinfection by different strains of APEC in the same animal with colisepticemia, the great antimicrobial resistance of these bacterial isolates and the genetic traits that modulate the virulence for high mortality rate of chickens for human consumption.

Toxoplasma gondii infection in dairy ewes: Vertical transmission and influence on milk production and reproductive performance.

The present study aimed to evaluate the transmission of toxoplasmosis (vertical and venereal) and its influence on milk production and reproductive problems of Lacaune sheep seropositives for Toxoplasma gondii. Males and females were serologically selected using indirect immunofluorescence method in three steps of the study. Step 1: In order to evaluate the influence of toxoplasmosis on milk production, the volume of milk produced by 40 sheep (22 seronegatives and 18 seropositives for T. gondii) was weekly measured throughout the lactation period. There were no significant differences between these two groups; in other words, toxoplasmosis did not affect milk production. Step 2: In order to assess T. gondii venereal transmission, five samples of semen from seropositive rams (n = 5) were tested by endpoint and real time PCR with two days of interval; however, these semen samples were PCR negatives for T. gondii. Step 3: To evaluate reproductive problems, 12 seropositive animals out of a flock of 68 pregnant ewes showed signs of reproductive problems, such as abortion or fetal resorption. T. gondii transplacental transmission was evaluated on blood drawn from newborn lambs (n = 41), and their respective seropositive mothers (n = 30) after single, double or triple births. Serological tests showed that 65.8% of the lambs had antibodies against this protozoan, indicating a high transmission from ewe to fetus during pregnancy. Therefore, it is concluded that toxoplasmosis in sheep may impair reproduction with a high percentage of vertical transmission.

Parasites in dairy cattle farms in southern Brazil / Los parásitos en las granjas de ganado lechero en el sur de Brasil

ABSTRACT Objective. The aim of this study was to investigate the relation among type and size of the farms related to the management of production and especially the presence and control of parasites. Materials and methods. To accomplish the above, 65 farms were used: 56 of semi-extensive system (cows in pastures), classified as low density farms (n=15, up to 15/cows), medium density farms (n=20; between 16-30/cows) and large density farms (n=21; over 31/cows). It was also included nine farms freestall system (n=9; confinement, over 70 cows) as part of the study population. Results. In the farms visited for the study can be noticed that the farms that had the higher level of technological improvement in managements practice were properties in high level (3) and freestall (4). In most of the farms there were observed the presence of ticks and flies, regardless of density of farms. Feces samples collected from 650 cows (n=10/farm), and analyzed using McMaster showed that 191 cows were positive for parasites. The presence of antibodies in bulk milk tank occurred in 36 (55.38%; CI95% 42-67) to Neospora caninum of the studied farms. Conclusions. We also conclude that N. caninum is present in cattle herds in Western Santa Catarina, and may be linked to reproductive problems in cows.

RESUMEN Objetivo. El objetivo de este estudio fue investigar la relación entre el tipo y el tamaño de las fincas relacionadas con la gestión de la producción, y especialmente, la presencia y el control de los parásitos. Materiales y métodos. Para lograr lo anterior, se utilizaron 65 granjas: 56 de semi-extensivo sistema (vacas en los pastos), clasificado como granjas de baja densidad (n=15, hasta 15/vacas), las granjas de densidad media (n=20; entre 16-30/vacas) y granjas grandes densidad (n=21; más de 31/vacas). También se incluyeron nueve del sistema de granjas freestall (n=9; confinamiento, más de 70 vacas) como parte de la población de estudio. Resultados. En las granjas visitadas se pudó notar que tenían el mayor nivel de mejora tecnológica en la práctica de gestiones eran propiedades en alto nivel (3) y estabulación libre (4). En la mayoría de las granjas no se observó la presencia de garrapatas y moscas, independientemente de la densidad de las explotaciones. Muestras de heces recogidas de 650 vacas (n = 10/granja), y analizados usando McMaster mostró que 191 vacas fueron positivas para parásitos. La presencia de anticuerpos en tanque de leche a granel se produjo en 36 (55.38%; IC95% 42-67) por Neospora caninum de las fincas estudiadas. Conclusiones. Se concluye que N. caninum está presente en los rebaños de ganado en el oeste de Santa Catarina, y puede estar vinculado a problemas reproductivos en vacas.

Diphenyl diselenide supplementation in infected mice by Toxoplasma gondii: Protective effect on behavior, neuromodulation and oxidative stress caused by disease.

The aim of this study was to evaluate the effect of subcutaneous administration of diphenyl diselenide (PhSe)2 on animal behavior and activities of acetylcholinesterase (AChE), adenylate kinase (AK), and creatine kinase (CK) in the brain of mice infected by Toxoplasma gondii. In addition, thiobarbituric acid reactive species (TBARS) levels and glutathione (GR, GPx and GST) activity were also evaluated. For the study, 40 female mice were divided into four groups of 10 animals each: group A (uninfected and untreated), group B (uninfected and treated with (PhSe)2), group C (infected and untreated) and group D (infected and treated with (PhSe)2). The mice were inoculated with 50 cysts of the ME49 strain of T. gondii. After infection the animals of the groups B and D were treated on days 1 and 20 post-infection (PI) with 5.0 µmol/kg of (PhSe)2 subcutaneously. Behavioral tests were conducted on days 29 PI to assess memory loss (object recognition), anxiety (elevated plus maze), locomotor and exploratory activity (Open Field) and it was found out that infected and untreated animals (group C) had developed anxiety and memory impairment, and the (PhSe)2 treatment did not reverse these behavioral changes on infected animals treated with (PhSe)2 (group D). The results showed an increase on AChE activity (P < 0.01) in the brain of infected and untreated animals (group C) compared to the uninfected and untreated animals (group A). The AK and CK activities decreased in infected and untreated animals (group C) compared to the uninfected and untreated animals (group A) (P < 0.01), however the (PhSe)2 treatment did not reverse these alterations. Infected and untreated animals (group C) showed increased TBARS levels and GR activity, and decreased GPx and GST activities when compared to uninfected and untreated animals (group A). Infected animals treated with (PhSe)2 (group D) decreased TBARS levels and GR activity, while increased GST activity when compared to infected and untreated animals (group C). It was concluded that (PhSe)2 showed antioxidant activity, but the dose used had no anti-inflammatory effect and failed to reverse the behavioral changes caused by the parasite.

Synergistic effects of resveratrol (free and inclusion complex) and sulfamethoxazole-trimetropim treatment on pathology, oxidant/antioxidant status and behavior of mice infected with Toxoplasma gondii.

This study aimed to investigate the synergistic effects of resveratrol and sulfamethoxazole-trimethoprim (ST) on the treatment of mice experimentally infected by Toxoplasma gondii during the chronic phase of the disease considering infection, behavior, and oxidative/antioxidants profile aspects. For the study, 60 mice were initially divided into two groups: uninfected (n = 24) and infected by T. gondii (n = 36). These two groups were later subdivided into other groups and treated with resveratrol (free and inclusion complex containing resveratrol) alone and co-administered with ST: groups A to D were composed by healthy mice and groups E to J were consisted of animals infected by T. gondii (VEG strain). Treatments began 20 days post-infection for 10 consecutive days with oral doses of 0.5 mg kg(-1) of ST (groups B and F), 100 mg kg(-1) of free resveratrol (groups C and G) and inclusion complex of resveratrol (nanoparticles containing resveratrol) (groups D and H), and lastly an co-administration of both drugs (groups I and J). Behavioral tests (memory, anxiety and locomotion) were performed after treatment. Liver and brain fragments were collected to evaluate pathological changes, brain cysts counts, as well as oxidant and antioxidant levels. A reduction on the number of cysts in the brain of animals treated with both drugs combined was observed; there was also reduced number of lesions on both organs. This drug combined effect was also able to reduce oxidative and increase antioxidant levels in infected mice, which might be interpreted as a resveratrol protective effect. In addition, the combination of ST and resveratrol was able to prevent behavioral changes in infected mice. Therefore, the use of co-administration drugs enhances the therapeutic effect acting on a synergic way, reducing the oxidizing effects of the chemical treatment for toxoplasmosis. In addition, resveratrol in inclusion complex when co-administered with ST showed an improved therapeutic effect of ST reducing oxidative damage, liver damage and the number of cysts in the brain of T. gondii infected mice.

Toxoplasma gondii: Effects of diphenyl diselenide in experimental toxoplasmosis on biomarkers of cardiac function.

This study aimed to investigate the effects of diphenyl diselenide (PhSe)2 to treat mice experimentally infected by Toxoplasma gondii on seric biomarkers of cardiac function (creatine kinase, creatine kinase MB, troponin, and myoglobin), and lactate dehydrogenase, as well as to evaluate the enzymatic activity of creatine kinase (CK) and adenylate kinase (AK) in heart tissue. For the study, 40 female mice were divided into four groups of 10 animals each: the group A (uninfected and untreated), the group B (uninfected and treated), the group C (infected and untreated) and the group D (infected and treated). The inoculation was performed with 50 cysts of T. gondii (ME-49 strain). Mice from groups B and D were treated at days 1 and 20 post-infection (PI) with 5 µmol kg(-1) of (PhSe)2 subcutaneously. On day 30 PI, the mice were anesthetized and euthanized for blood and heart collection. As a result, it was observed a decrease in AK activity (P < 0.01) in the heart samples of groups C and D compared to the group A. Cardiac CK increased in the group C compared to the group A (P < 0.01). CK levels increased in infected mice (the group C) compared to other groups (A and D). Regarding CK-MB level, there was a decrease in the group D compared to the group B, without statistical difference compared to control groups (A and C). It was observed an increase on myoglobin in groups C and D, differently of troponin, which did not show statistical difference (P < 0.05) between groups. Mice from the group C showed an increase in lactate dehydrogenase (LDH) levels compared to other groups (A, B, and D). Histopathological evaluation of heart samples revealed necrosis, hemorrhagic regions and inflammatory infiltrates in mice from the Group C, differently from the group D where animals showed only inflammatory infiltrates. Based on these results we conclude that the (PhSe)2 had a protective effect on the heart in experimental toxoplasmosis by modulating tissue and seric CK activity, and avoiding an increase on seric LDH levels, probably due to the antioxidant effect of this compound.

Canine cutaneous neosporosis in Brazil.

The clinical signs of infection in dogs with Neospora caninum are usually associated with neurological disorders and are seen in young dogs. In this brief case report we observed multifocal ulcerative and exudative skin nodules on the neck and pelvic limbs of a 10-year-old cocker spaniel dog. Infection with N. caninum was diagnosed on the basis of cytology and examination of skin tissues by PCR. The dog initially responded to treatment with clindamycin and then relapsed; the dog died. Infection with N. caninum may have been due in part to immune suppression due to hyperadrenocorticism; which either allowed for the development of a primary infection or reactivation of a latent infection by N. caninum with the occurrence of skin lesions.

Effects of sulfamethoxazole-trimethoprim associated to resveratrol on its free form and complexed with 2-hydroxypropyl-ß-cyclodextrin on cytokines levels of mice infected by Toxoplasma gondii.

The aim of this study was to investigate the effects of resveratrol on its free form and complexed with 2-hydroxypropyl-ß-cyclodextrin (HPßCD) when associated with sulfamethoxazole-trimethoprim (ST) on cytokines levels of mice (n = 60) experimentally infected by Toxoplasma gondii. Groups A and E were used as controls (untreated): negative and positive, respectively. The onset of treatment started 20 days post-infection (PI), and it lasted for 10 consecutive days. ST was administered orally in doses of 0.5 mg kg(-1) for groups B and F, while 100 mg kg(-1) was the dose for resveratrol in its free form (groups C - G), inclusion complex (groups D and H), and on free and inclusion complex together (groups I - J). On day 31 PI, blood samples were collected in order to evaluate the cytokine profile. The mice that received drug combination (I and J) showed a significant (P < 0.05) reduction in the number of cysts in the brain compared to other infected groups (E - H). The results showed that mice from the Group E had increased (P < 0.001) levels of pro-inflammatory cytokines, while IL-10 levels were reduced when compared to the Group A. Additionally, there were increased levels of IL-4 and IFN-γ in animals of groups C and D, respectively (P < 0.05). Animals of the Group B showed reduced levels of IL-1, IL-4, IL-6, TNF-α, and IFN-γ (P < 0.05). Mice infected and treated (groups F - J) showed increased levels of pro-inflammatory cytokines along with a reduction of IL-10. Treatment with the combination of drugs (the Group J) led to a protective effect, i.e. reduction in pro-inflammatory cytokines. Therefore, resveratrol associated with ST was able to modulate seric cytokine profile and moderate the tissue inflammatory process caused by T. gondii infection, as well as to reduce parasite multiplication.

Sulfamethoxazole-trimethoprim associated with resveratrol for the treatment of toxoplasmosis in mice: Influence on the activity of enzymes involved in brain neurotransmission.

This study aimed to investigate the influence of sulfamethoxazole-trimethoprim (ST) associated with resveratrol on the enzymatic activities of acetylcholinesterase (AChE), adenylate kinase (AK), pyruvate kinase (PK), and creatine kinase (CK) in the brain of mice experimentally infected by Toxoplasma gondii. For that, 60 mice were divided into ten groups with 6 animals each: groups A to D composed by healthy mice and groups E to J consisting of animals infected by T. gondii (VEG strain). Animals started treatment 20 days post-infection for 10 consecutive days with oral doses of 0.5 mg kg(-1) of ST (groups B and F), 100 mg kg(-1) of free resveratrol (groups C and G) and inclusion complex of resveratrol (nanoparticles containing resveratrol) (groups D and H), as well as with an association of both drugs (groups I and J). The results showed increased (P < 0.001) AChE activity on infected animals (groups E-J) when compared to not-infected (A) animals, and also uninfected animals treated with ST (group B) had increased AChE activity. AK activity decreased (P < 0.001) in the infected and untreated (group E), differently from the other groups that did not differ. PK activity did not differ between groups (P > 0.05). When comparing control groups (uninfected (A) and infected (E)), we verified a significant (P < 0.001) increase in CK activity in the brain, and it is noteworthy that the animals treated with resveratrol associated with ST (group I and J) had similar CK activity to those animals from the group A. Treatment with the combination of ST and resveratrol was able to reduce (P < 0.05) the number of parasitic cysts in the brain, thus reduced inflammatory infiltrates in the liver, and prevented the occurrence of hepatocytes lesions due to toxoplasmosis in mice. Based on these results, it is possible to conclude that increased AChE and CK activities after T. gondii infection did not change with the treatment of ST-resveratrol association. In addition, decreased AK activity caused by T. gondii infection was normalized by ST-resveratrol treatment. T. gondii infection and treatment does not affect PK activity in brain.

Biochemical detection of E-ADA on Neospora caninum tachyzoites and the effects of a specific enzymatic inhibitor / Detección bioquímica de E-ADA en taquizoitos de Neospora caninum y los efectos de un inhibidor enzimático específico

Objective. This study aimed to investigate the presence and activity of the ecto adenosine deaminase (E-ADA) enzyme in tachyzoites of Neospora caninum (Nc-1 strain), as well as to assess the activity of a well-known E-ADA inhibitor, the deoxycoformycin. Materials and methods. The parasites were grown in cell culture, being subsequently separated in a pellet of tachyzoites, on which the E-ADA activity was tested using the concentrations 0 (control), 0.2, 0.4 and 0.8 mg mL-1. Results. The E-ADA showed high activity, progressively increasing its activity according to the enhancement of the protein concentration. The test was carried out with different concentrations of deoxycoformycin, showing that it was able to inhibit the E-ADA present on the free form of the parasite. Conclusions. Based on these results we conclude that the E-ADA is present on tachyzoites of N. caninum, and deoxycoformycin is able to inhibit this enzyme. In this sense, knowing the negative impact of N. caninum on reproductive issue in cattle (mainly abortion), might it is an alternative in order to deal with this parasitic infection.

Objetivo. Este estudio tuvo como objetivo investigar la presencia y la actividad de la adenosina deaminasa (E-ADA) en taquizoitos de Neospora caninum (Nc-1 cepa), así como ensayar un conocido inhibidor de estas enzimas. Material y métodos. Los parásitos se multiplicaron en cultivo celular y posteriormente fueron separados en un sedimento de taquizoitos. La actividad de E-ADA fue probada en el parásito, usando concentraciones de 0 (control), 0.2, 0.4 y 0.8 mg mL-1. Resultados. El E-ADA tenía una alta actividad aumentó progresivamente de acuerdo con la concentración de proteínas. La prueba se llevó a cabo con diferentes concentraciones del inhibidor E-ADA (desoxicoformicina) y fue capaz de inhibir la presente E-ADA en el parásito. Conclusiones. Por la observación de los resultados se concluye que la enzima E-ADA está presente en taquizoítos de N. caninum, y desoxicoformicina es capaz de inhibir la acción de la enzima. En este sentido, conocer el impacto negativo del parásito en los problemas productivos y reproductivos en el ganado (principalmente abortos), estos resultados pueden ser una alternativa para evitar esta enfermedad parasitaria.

Seroprevalence and risk factors for Neospora caninum in goats in Santa Catarina state, Brazil.

Neosporosis is an infectious disease caused by the parasite Neospora caninum. Knowledge regarding neosporosis in goats is still quite limited, especially in the state of Santa Catarina (SC), southern Brazil. Therefore, this study aimed to assess the seroprevalence and risk factors for N. caninum in goats in the western and mountain regions of SC. Blood samples were collected from 654 goats in 57 municipalities. The indirect immunofluorescence test was used for antibody detection against N. caninum. Thirty samples (4.58%) were seropositive, with titers ranging from 1:50 to 1:6400. An epidemiological survey was also conducted in order to identify risk factors for neosporosis in goats. It was found that reproductive problems on the farms, as well as the diet and direct contact with dogs were casual risks for neosporosis. These results indicate that N. caninum infects goats in these regions, which may lead to reproductive problems.