Activity of cumin essential oil to control fascioliasis: Efficacy and changes in the tegument of Fasciola hepatica.

Fasciola hepatica, the liver trematode, infects ruminants and causes economic loss. Because parasites are developing resistance to commercial drugs, the negative effects of parasitism are increasing. In this study, we aimed to evaluate the efficacy of cumin (Cuminum cyminum) essential oil against F. hepatica eggs and adults. The eggs were incubated with eight concentrations of the essential oil (0.031125-4.15 mg/mL), and viable eggs were counted after 14 days and classified as embryonated or non-embryonated. Adult flukes were incubated in Roswell Park Memorial Institute medium to ensure their viability and then incubated in essential oil. They were observed for 24 h after treatment. The adults were assessed with the two lowest effective oil concentrations used in the ovicidal test. Three controls were used for both tests: nitroxynil, a negative control, and Tween®80. After incubation in oil, the adult specimens were processed for histological analysis and stained with hematoxylin-eosin. In addition, the oil was tested for cytotoxicity using Madin-Darby bovine kidney cells to assess any possible effect on them. The oil was effective in ovicidal and adulticidal inhibition of the trematode, with statistically significant results. All concentrations assessed in the ovicidal test were 100% effective. The adult test was effective within 15 h and inactivated all the specimens at the highest concentration evaluated (0.06225 mg/mL). Histological analysis showed that cumin essential oil resulted in marked areas of vacuolization. The spines showed no structural changes but were surrounded by microvesicles. These findings indicated that cumin oil could be a potential compound in the control of fasciolosis.

Microscopic alterations in Fasciola hepatica treated with the essential oils of Pelargonium graveolens and Citrus aurantium.

Fasciolosis is one of the most important parasitic diseases in animal health, affecting mainly ruminants, causing economic and productivity losses. This study aimed to evaluate the in vitro ovicidal and adulticidal activity of essential oils (EOs) from Pelargonium graveolens (geranium) and Citrus aurantium (sour orange) on Fasciola hepatica. Performed Gas Chromatography of EOs P. graveolens and C. aurantium, with major compound, citronellol (31.37 %) and limonene (93.89 %), respectively. For the cytotoxicity assay, the sour orange EO showed to be promising when used in lower concentrations. For the ovicidal tests, the eggs were incubated with geranium EOs at concentrations from 4.5 to 0.03375 mg/mL and sour orange at concentrations from 4.25 to 0.031875 mg/mL, along with controls. The viable eggs were counted on the 14th day post-incubation. Adult forms of F. hepatica were incubated containing the EOs and observed for 24 h after treatment, as well as the control groups. Later the specimens were fixed for histological analysis. Geranium and sour orange EOs in trematode eggs at the concentrations tested were 100 % effective in inactivating hatching (p < 0.05) when compared to the untreated control. In the adulticidal test, the essential oil of P. graveolens at both concentrations tested (0.0675 and 0.03375) within 15 h, promoted the death of flukes. For C. aurantium, 18 h was enough to inactivate all specimens, up to a concentration of 0.06375. The histological analysis, observed the accumulation of liquid in the tegument in the specimens incubated in C. aurantium and P. graveolens, with vacuolization in the tegument and spines, preventing externalization. The results of the study present OEs with efficient ovicidal and adulticidal activity.

Gastrointestinal Parasites in Sheep from the Brazilian Pampa Biome: Prevalence and Associated Factors.

Parasitic infections are common in sheep farming worldwide. A cross-sectional study was carried out to determine the prevalence and associated risk factors among gastrointestinal parasites and sheep herds from the Brazilian Pampa biome, Rio Grande do Sul state, Brazil. Twenty-one farms were visited, where rectal faecal samples were individually collected from 309 animals. The sheep industry on the studied farms was characterised by small farms with a low level of technification, an extensive grazing system and meat production. Of all samples analysed, strongyle nematodes had the highest prevalence (77.02%), followed by Eimeria spp. (70.55%), Moniezia expansa (20.39%) and Strongyloides papillosus (17.48%). Mixed infection, between helminths and protozoans, was detected in 68.61% of samples. All herds were positive for strongyle and Eimeria spp. A predominance of Haemonchus spp. and Trichostrongylus spp. nematodes was observed in the herds. Younger animals were significantly more affected by Eimeria and M. expansa. In the semi-intensive and intensive systems, a higher frequency of Eimeria and strongyle infections was observed. Parasite infection was significantly reduced at low animal densities. The Brazilian Pampa region presents a high prevalence of gastrointestinal parasites among sheep; age, breeding system and stocking density were factors associated with parasite infection.

As infecções parasitárias são comuns na criação de ovinos em todo o mundo. Um estudo transversal foi realizado para determinar a prevalência e fatores de risco associados entre parasitoses gastrintestinais e rebanhos ovinos do bioma Pampa brasileiro, estado do Rio Grande do Sul, Brasil. Vinte e uma fazendas foram visitadas, onde foram coletadas amostras de fezes individuais da ampola retal de 309 animais. A produção ovina era caracterizada por pequenas propriedades com baixo nível de tecnificação, os animais eram criados em sistema extensivo e para produção de carne. De todas as amostras analisadas, os estrongilídeos (Nematoda: Strongylida) foram mais prevalentes (77,02%), seguido por Eimeria spp. (70,55%), Moniezia expansa (20,39%) e Strongyloides papillosus (17,48%). Infecção mista, entre helmintos e protozoários, foi detectada em 68,61% das amostras. Todos os rebanhos foram positivos para estrongilídeos e Eimeria spp. Foi observada predominância dos nematódeos Haemonchus spp. e Trichostrongylus nos rebanhos. Animais mais jovens foram significativamente mais afetados por Eimeria e M. expansa. Nos sistemas semi-intensivo e intensivo, observou-se maior frequência de infecções por Eimeria e estrongilídeos. A infecção parasitária foi significativamente menor em fazendas com baixa densidade animal. A região do Pampa brasileiro apresenta alta prevalência de parasitos gastrintestinais em ovinos; idade, sistema reprodutivo e densidade animal foram fatores associados à infecção parasitária.

The relation between columbiform birds (Columba livia) age and gender and infection indices of rhinonyssid and ereynetid mites.

Respiratory systems of birds may be parasitized by rhinonyssid and ereynetid mites, which are commonly found in their nasal cavities. Levels of infection caused by mites in relation to maturity and gender of birds have been poorly investigated. Although Columba livia is the host of both Tinaminyssus and Trispeleognathus species worldwide, there are no data on prevalence and mean intensity of infection by mites in immature and adult males and females. Therefore, this study aims to analyze infections in relation to characteristics of hosts in southern Brazil. In the sample of 160 birds under study, 24.38% were parasitized by rhinonyssid mites (Tinaminyssus melloi and Tinaminyssus columbae) while 5.0% were infested with ereynetid ones (Trispeleognathus striatus). Infections by rhinonyssid mites in immature and adult birds, as well as in male and female ones, were similar and there were no significant differences in prevalence and mean intensity of infection. However, when infections were analyzed separately, prevalence of T. melloi was significantly higher in immature (19.77%) than in adult (6.76%) birds (p = 0.021). Ereynetid mites, which parasitized only males, were more prevalent in adult (9.46%) than in immature (1.16%) birds (p = 0.025). Reproductive and behavioral characteristics of C. livia (e.g., year-round breeding and social behavior) may enable transmission and maintenance of mite populations, since direct contact among hosts and the presence of mites in their nests are considered forms of transmission. Even though information on the life history of mites is important to understand parasite-host relations, there is a gap in the biology of species commonly found in C. livia, which constitutes an area to be explored.

Cryptosporidium spp. in Columba livia Gmelin, 1789 (Columbiformes: Columbidae) free-living pigeons from urban areas in Pelotas, Rio Grande do Sul, Brazil.

Cryptosporidium spp. are parasites with zoonotic potential that cause intestinal diseases, generally intense diarrheal, on their hosts, which tend to be immunocompromised. Large populations of pigeons in urban environments can lead to greater human exposure to Cryptosporidium spp., as this bird is considered a potential reservoir and is able to transmit several pathogens. This study aimed in determining the occurrence of Cryptosporidium spp. oocysts in feces of free-living pigeons (Columba livia) found in urban areas in the city of Pelotas, Rio Grande do Sul, south of Brazil. Fecal samples (n = 50) were collected from young and adult pigeons captured in different locations in the urban area and the parasitological diagnosis was performed through Ritchie's modified technique and Kinyoun's technique. Among the 50 samples, 18% (IC95% 9.7-30.8) were positive for Cryptosporidium spp. with a low number of oocysts being detected on fecal smears. Our results confirmed the occurrence of Cryptosporidium spp. oocysts in feces of free-living pigeons from the urban area of the city of Pelotas. This is the first report of Cryptosporidium spp. oocysts in feces of pigeons in south Brazil. This data has epidemiological significance because the oocysts could be from zoonotic species, which consequently shows that humans might be exposed to infection.

Serum anti-Toxoplasma gondii antibodies in Passer domesticus (Linnaeus, 1758) (Passeriformes: Passeridae), in the municipality of Pelotas, RS, Brazil / Anticorpos séricos anti-Toxoplasma gondii em pardais, Passer domesticus (Linnaeus, 1758) (Passeriformes: Passeridae), do município de Pelotas, RS, Brasil

The objective of this study was to detect the presence of anti-Toxoplasma gondii antibodies in serum samples from 100 house sparrows (Passer domesticus Linnaeus, 1758) that were caught in the municipality of Pelotas, Rio Grande do Sul, Brazil. The modified agglutination test (MAT) was used to investigate anti-T. gondii antibodies and samples with a cut-off dilution > 5 were considered positive. Among the 100 serum samples analyzed, 80 (80%) were reactive. These results demonstrate that P. domesticus may play an important role in the epidemiological chain of T. gondii, since it is widely distributed throughout Brazil, and may act as a source of infection to domestic and wild felids.(AU)

O objetivo deste estudo foi detectar a presença de anticorpos anti-Toxoplasma gondii em amostras de soro de 100 pardais (Passer domesticus Linnaeus, 1758) capturados na área urbana do município de Pelotas, Rio Grande do Sul, Brasil. Para a pesquisa de anticorpos anti-T. gondii foi utilizado o teste de aglutinação modificado (MAT) e foram consideradas positivas as amostras que apresentaram título > 5. Das 100 amostras de soro analisadas, 80 (80%) foram reagentes. Esses resultados demonstram que P. domesticus, por ser amplamente distribuído em todo país, pode desempenhar um papel importante na cadeia epidemiológica de T. gondii, podendo atuar como fonte de infecção para felinos domésticos e silvestres.(AU)

An indirect ELISA for Neosporosis: Associating recombinant Neospora caninum proteins NcSRS2 and NcSAG1.

Neosporosis is caused by infection with the protozoa Neospora caninum. It manifests as various neurological symptoms and is considered as one of the main causes of abortion in cattle, and induces uncommon congenital infection in sheep. The standard diagnosis is based on indirect immunofluorescence (IFI); however, cross-reactivity with other protozoa proteins is common. Aiming a more specific diagnosis, recombinant antigens have been tested in several immunoassays; of these, NcSAG1 (surface antigen-1) and NcSRS2 (SAG1-related sequence 2) were the most promising. In this context, we developed an indirect ELISA with recombinant NcSRS2 (ELISA-rNcSRS2) and NcSAG1 (ELISA-rNcSAG1) proteins alone and in association (ELISA-rNcSRS2/rNcSAG1) for the diagnosis of cattle and ovine neosporosis. A total of 216 samples from cattle and 154 samples from sheep were used to evaluate the ELISAs. The sensitivity and specificity results of the ELISA-rNcSRS2 were 91.5 % and 96.4 % for cattle, and 89.6 % and 96.3 % for sheep, respectively. For the ELISA-rNcSAG1, the sensitivity and specificity were 84.9 % and 97.3 % for cattle, and 89.6 % and 92.6 % for sheep, respectively. The sensitivity and specificity of the ELISA-rNcSRS2/rNcSAG1 was 98.1 % and 99.1 % for cattle, 100 % and 97.2 % for sheep, respectively. These results indicated that indirect ELISA using the rNcSRS2 and rNcSAG1 proteins are a highly sensitive and specific method, especially when used in association, for detecting antibodies in cattle and ovine populations infected with N. caninum.

The raptor Chimango Caracara (Milvago chimango) (Aves: Falconiformes) - A new host for Trichomonas gallinae (protozoa: Trichomonadidae).

This work describes a specimen of Chimango Caracara (Milvago chimango - Vieillot, 1816), from southern Brazil, as a new natural host for Trichomonas gallinae (Rivolta, 1878). Caseous oral lesions were observed in a young bird, and the parasite was isolated in modified Diamond's media. Morphology of the parasite was evaluated through microscopy and subsequently, sequencing of the internal transcribed spacer 1 (ITS1) of ribosomal DNA was performed to confirm T. gallinae identification. As far as authors are concerned, this is the first report of Milvago chimango as a natural host for T. gallinae.

Oral trichomoniasis in raptors in Southern Brazil / Tricomoníase oral em rapinantes no sul do Brasil

A retrospective study of oral trichomoniasis cases in raptors was carried out at "Laboratório Regional de Diagnóstico" of "Faculdade de Veterinária" of "Universidade Federal de Pelotas" (LRD-UFPel) from December 2014 to August 2017. Seven necropsy reports were reviewed. All raptors including Falconiformes and Strigiformes orders were from periurban zones. Four birds were adults, and there was no apparent sex predisposition. Clinical signs included dysphagia, regurgitation, and anorexia. Gross lesions were characterized by multifocal yellow to coalescent caseous nodules adhered to the oral cavity, which extended to the entrance of the esophagus. Microscopically, the lesions were characterized by severe focally extensive heterophilic granulomatous stomatitis. Trichomonas gallinae was isolated in modified Diamond medium of all samples collected from birds. It is believed the transmission occurred by the predation of domestic pigeons (Columbia livia domestica) contaminated with T. gallinae that agglomerate in patios of grain processing complexes.(AU)

Foi realizado um estudo retrospectivo de casos de tricomoníase oral em rapinantes recebidos no Laboratório Regional de Diagnóstico da Faculdade de Veterinária da Universidade Federal de Pelotas (LRD-UFPel) no período de dezembro de 2014 a agosto de 2017. Foram revisados os protocolos de necropsia encontrando-se sete casos. Os animais eram todos de zonas periurbanas e foram afetadas a ordem Falconiformes e a ordem Strigiformes. Das aves afetadas 4/7 eram adultas e não houve aparente predisposição por sexo. Os sinais clínicos incluíram disfagia, regurgitação e anorexia. As lesões se caracterizaram por nódulos amarelados multifocais a coalescentes aderidos a cavidade oral, que se estendiam até a entrada do esôfago. Histologicamente as lesões caracterizaram-se por estomatite heterofílica granulomatosa focalmente extensiva acentuada. Foi isolado em meio Diamond modificado Trichomonas gallinae de todas as amostras coletadas das aves. Acredita-se que a fonte de transmissão tenha ocorrido a partir da predação por estes rapinantes de pombas domésticas (Columbia livia domestica) portadoras com T. gallinae que se aglomeram em pátios de complexos de beneficiamento de grãos.(AU)

Trichomonas vaginalis and HIV infection: relation and prevalence among women in southern Brazil

Trichomoniasis is a curable sexually transmitted infection (STI) that has been reported to be linked to exposure to human immunodeficiency virus (HIV), although few studies have described this association. The purpose of this study was to focus on the incidence of trichomoniasis in low-income women, its relation to HIV status, viral load levels and TCD4+cell counts, among other risk factors, using an in vitro culture as a diagnostic test. A cross-sectional study among 267 women (103 HIV-positive and 164 HIV-negative) was conducted in 2015. The overall prevalence of Trichomonas vaginalis (TV) infection was 6.4%. Among HIV-positive and HIV-negative women, the prevalence was 3.9% and 7.9%, respectively, yet these results were not statistically different (p=0.1878). The factors associated with TV infection were cigarette smoking (OR= 3.52), vaginal itching (OR=4.43) and bacterial vaginosis (BV) (OR= 5.29). HIV status, TCD4+ cell count and viral load were not associated with TV infection in this group. The prevalence rates found, lower than those observed in other studies, may be due to the fact that the women evaluated in the present study are part of a low-risk population as well as the limited sample size of HIV positive women

Aptima Trichomonas vaginalis assay elucidates significant underdiagnosis of trichomoniasis among women in Brazil according to an observational study.

OBJECTIVES: Trichomonas vaginalis (TV) infection is the most common non-viral STI globally and can result in adverse pregnancy outcomes and exacerbated HIV acquisition/transmission. Nucleic acid amplification tests (NAATs) are the most sensitive diagnostic tests, with high specificity, but TV NAATs are rarely used in Brazil. We investigated the TV prevalence and compared the performance of the US Food and Drug Association-cleared Aptima TV assay with microscopy (wet mount and Gram-stained) and culture for TV detection in women in Pelotas, Brazil in an observational study. METHODS: From August 2015 to December 2016, 499 consecutive asymptomatic and symptomatic sexually active women attending a Gynaecology and Obstetrics Outpatient Clinic were enrolled. Vaginal fluid and swab specimens were collected and wet mount microscopy, Gram-stained microscopy, culture and the Aptima TV assay performed. RESULTS: The median age of enrolled women was 36.5 years (range: 15-77). The majority were white, had a steady sexual partner and low levels of education. The TV detection rate was 4.2%, 2.4%, 1.2% and 0% using the Aptima TV assay, culture, wet mount microscopy and Gram-stained microscopy, respectively. The sensitivity of culture and wet mount microscopy was only 57.1% (95% CI 36.5 to 75.5) and 28.6% (95% CI 13.8 to 50.0), respectively. CONCLUSIONS: A 4.2% positivity rate of T. vaginalis was found among women in Pelotas, Brazil and the routine diagnostic test (wet mount microscopy) and culture had low sensitivities. More sensitive diagnostic tests (NAATs) and enhanced testing of symptomatic and asymptomatic at-risk women are crucial to mitigate the transmission of TV infection, TV-associated sequelae and enhanced HIV acquisition and transmission.

Rhinonyssidae (Acari) in the house sparrows, Passer domesticus (Linnaeus, 1758) (Passeriformes: Passeridae), from southern Brazil / Ácaros nasais Rhinonyssidae parasitos de Passer domesticus (Linnaeus, 1758) (Passeriformes: Passeridae) no extremo sul do Brasil

Abstract We report the occurrence and infection parameters of two species of nasal mites in Passer domesticus (Linnaeus, 1758) (house sparrow). Nasal passages, trachea, lungs, and air sacs of 100 house sparrows captured in an urban area at the city of Pelotas, State of Rio Grande do Sul, southern Brazil, were examined with a stereomicroscope. The mite, Sternostoma tracheacolum Lawrence, 1948 was present in the trachea and/or lungs (or both) of 13 birds (13%) at a mean intensity of 6.7 mites/infected host. Ptilonyssus hirsti (Castro & Pereira, 1947) was found in the nasal cavity of 1 sparrow (1%); coinfection was not observed in this bird. There was no significant difference in the prevalence and mean intensity of infection of S. tracheacolum between male and female birds. To our knowledge, this is the first study reporting the occurrence of S. tracheacolum in P. domesticus in Brazil and the presence of P. hirsti in P. domesticus from Rio Grande do Sul, Brazil. This is the first survey to provide the infection parameters of each of these mites in house sparrows.

Resumo O objetivo deste estudo foi relatar duas espécies de ácaros nasais parasitos do sistema respiratório de Passer domesticus (Linnaeus, 1758) (pardal) e seus respectivos índices parasitológicos. Para isso, foram examinados ao estereomicroscópio, cavidade nasal, traqueia, pulmão e sacos aéreos de 100 pardais capturados na área urbana de Pelotas, Rio Grande do Sul, Brasil. Sternostoma tracheacolum Lawrence, 1948 ocorreu na traqueia e/ou pulmão de 13 aves (13%) com intensidade média de 6,7 ácaros/hospedeiro infectado e Ptilonyssus hirsti (Castro & Pereira, 1947) na cavidade nasal de um único pardal (1%), não havendo coinfecção. A prevalência e intensidade média de infecção por S. tracheacolum entre hospedeiros machos e fêmeas não apresentou diferença significativa. Este relato caracteriza o primeiro registro de S. tracheacolum parasitando P. domesticus no Brasil e de P. hirsti em pardais no RS, Brasil, e seus respectivos índices de infecção.

Occurrence of intestinal parasites in collectors of recyclable materials in southern Brazil

Solid waste produced by modern society, together with lack of basic sanitation and poor hygiene practices result in risks to public health. 46 workers from the cities of Pelotas, Canguçu, Morro Redondo, Rio Grande and Capão do Leão were evaluated by parasitological examination of two fecal samples by Ritchie, Hoffman, Pons and Janer- HPJ techniques and Sheather method as well as a consented interview. 47.8% (22/46) of the examined individuals were parasitized, with no significant difference between both groups (garbage dump and recycling volunteers). Most of the samples evaluated presented forms of helminths (68.2%) while protozoa were found in 31.8% of the samples. Helminths found, and their respective prevalence were: Ascaris lumbricoides (31.8%), Ascaris lumbricoides/Trichuris trichiura (18.2%), Trichuris trichiura (9.1%), Taenia sp (4.6%) and Trichuris trichiura/Ancylostomids (4.6%). Protozoa were: Giardia intestinalis (13, 6%), Entamoeba spp. (9.1%) and Endolimax nana (9.1%). Monoparasitism was observed in 77.3% of the cases. Most of the population consisted of women, 21-40 years of age, low educational level and monthly income less than one minimum wage. The majority of the workers did not use protective equipment. Multivariate analysis revealed that infection from parasites in scavengers who lived near open sewers was 6.65 times higher (p = 0.0065) than in the other workers. In the localities studied, our results showed that the risk of acquiring parasitic infections was related to poor sanitation and the lack of sewerage systems; and not actually due to handling these types of waste. The parasitological indices found are similar to those described previously in other parts of the country

Rhinonyssidae (Acari) in the house sparrows, Passer domesticus (Linnaeus, 1758) (Passeriformes: Passeridae), from southern Brazil.

We report the occurrence and infection parameters of two species of nasal mites in Passer domesticus (Linnaeus, 1758) (house sparrow). Nasal passages, trachea, lungs, and air sacs of 100 house sparrows captured in an urban area at the city of Pelotas, State of Rio Grande do Sul, southern Brazil, were examined with a stereomicroscope. The mite, Sternostoma tracheacolum Lawrence, 1948 was present in the trachea and/or lungs (or both) of 13 birds (13%) at a mean intensity of 6.7 mites/infected host. Ptilonyssus hirsti (Castro & Pereira, 1947) was found in the nasal cavity of 1 sparrow (1%); coinfection was not observed in this bird. There was no significant difference in the prevalence and mean intensity of infection of S. tracheacolum between male and female birds. To our knowledge, this is the first study reporting the occurrence of S. tracheacolum in P. domesticus in Brazil and the presence of P. hirsti in P. domesticus from Rio Grande do Sul, Brazil. This is the first survey to provide the infection parameters of each of these mites in house sparrows.

Seroprevalence of Toxoplasma gondii and Toxocara canis in a human rural population of Southern Rio Grande do Sul.

Due to the growing population of pets, especially homeless dogs and cats, zoonoses still represent a significant public health problem. Toxoplasma gondii and Toxocara spp. are epidemiologically important zoonotic agents as they are etiological factors of human toxoplasmosis and toxocariasis, respectively. These parasites remain neglected even though they are substantially prevalent in rural areas. The aim of this study was to investigate T. gondii and T. canis seroprevalence and risk factors of seropositivity in a rural population in Pelotas municipality, Brazil. The study participants (n=344) were patients of a Basic Healthcare Unit (BHU) located in Cerrito Alegre. Blood samples were collected and tested for T. gondii antibodies by indirect immunofluorescence and T. canis antibodies by an indirect ELISA that targets an excreted-secreted antigen (TES). T. gondii seropositivity was 53.2%, with higher titers (1:256 - 1:1,024) in individuals who habitually eat pork, beef, or chicken, while T. canis seropositivity was 71.8% and concomitant T. gondii and T. canis seropositivity was 38.3%. Among the seropositivity risk factors assessed, only habitual undercooked meat consumption was significant (p = 0.046; OR = 3.7) for T. gondii and none of them were associated with T. canis seropositivity. Both parasites have a high prevalence in rural areas, which reinforces the need to invest in rural community education and health.

Characterization of Toxoplasma gondii isolates from herds of sheep in southern Brazil reveals the archetypal type II genotype and new non-archetypal genotypes.

Recent studies have demonstrated that, in Brazil and South America, strains of Toxoplasma gondii are often genotypically and biologically different from those found in countries on other continents. The objective of this study was to genotypically characterize T. gondii isolates from naturally infected sheep in herds in the southern region of the state of Rio Grande do Sul, Brazil, by means of the polymerase chain reaction with restriction fragment length polymorphism (PCR-RFLP). Five T. gondii isolates obtained from sheep in five municipalities in the state of Rio Grande do Sul were used. Application of multilocus PCR-RFLP multilocus using 12 genetic markers (SAG1, 5'3' SAG2, alt. SAG2, SAG3, BTUB, c22-8, c29-2, GRA6, L358, PK1, APICO and CS3) revealed four different genotypes in the five isolates studied: clonal type II (TgOvBrRS4), type BrIV (TgOvBrRS2 and TgOvBrRS3) and two new non-archetypal genotypes, ToxoDB-RFLP#270 and #271 (TgOvBrRS1 and TgOvBrRS5, respectively). The genotype structure found in the T. gondii isolates from naturally infected sheep in the southern region of Brazil was revealed to have high diversity. This study confirms the presence of rare circulation of the clonal type II genotype in Brazil.

PCR-based identification of Neospora caninum in the umbilical cord of a newborn calf in Brazil / Identificação de Neospora caninum por PCR em cordão umbilical de bezerros recém nascido no Brasil

ABSTRACT: This study was conducted at a beef cattle breeding farm in the far southern region of Brazil. The birth of a calf with unilateral corneal opacity was immediately reported to the Laboratory of Parasitology, in the Universidade Federal de Pelotas, Rio Grande do Sul, Brazil; blood was collected from the cow and calf before colostrum intake. The umbilical cords from this calf and from six other healthy animals were collected. Serological examination, utilizing an indirect fluorescent antibody test, was done using a cut-off point of 1:100. Polymerase chain reaction (PCR) was also performed using the umbilical cord samples. Serological tests showed that the calf was positive for Neospora caninum at birth, with a titer of 1:1600; a titer of 1:3200 was reported in the dam. PCR, using umbilical cord tissue from the affected animal, was positive for the presence of this parasite, and the molecular identity of the amplified product was confirmed by sequencing. Therefore, the detection of N. caninum DNA in the umbilical cord represents a novel alternative test for the diagnosis of this parasitic infection in newborn calves that are clinically suspected to have neosporosis.

RESUMO: O estudo foi realizado em uma fazenda de criação de gado de corte na região do extremo sul do Brasil. O nascimento de um bezerro com opacidade unilateral da córnea foi imediatamente relatado ao Laboratório de Parasitologia da Universidade Federal de Pelotas (RS) e coletou-se sangue da vaca e do bezerro antes da ingestão do colostro. Foram coletados sangue e cordão umbilical do bezerro com opacidade na córnea bem como de outros seis bezerros aparentemente saudáveis antes da ingestão de colostro. Foram realizados exames sorológicos utilizando o teste indireto de anticorpos fluorescentes (IFAT) com ponto de corte de 1: 100 e reação em cadeia da polimerase (PCR). O resultado dos exames sorológicos mostraram que o bezerro foi positivo para N. caninum ao nascer com titulação de 1:1600 e o de sua mãe foi de 1:3200 na IFAT. A PCR do cordão umbilical foi positiva para o agente no animal com opacidade córnea unilateral e a identidade molecular do produto amplificado foi confirmada por sequenciamento. A detecção de DNA de N. caninum no cordão umbilical é uma nova alternativa para o diagnóstico deste agente em bezerros recém-nascidos com suspeita clínica de neosporose.

The incidence and productive significance of ovine toxoplasmosis in Southern Brazil / Incidência e significância econômica da toxoplasmose ovina no sul do Brasil

ABSTRACT: This research aimed to determine incidence of Toxoplama gondii infection in Corriedale sheep in southern Brazil during pregnancy and the extent of associated losses. Blood samples were collected from 411 Corriedale sheep in two flocks at different locations in the state of Rio Grande do Sul (Brazil). Presence of T. gondii was diagnosed by the Indirect Fluorescent Antibody Technique (IFAT). Seroprevalence of T. gondii during pre-mating was 20.2%, with no significant difference between the two flocks studied. Infection by T. gondii was influenced by ewes'age (P≤0.05). The pre-mating seroprevalence did not influence either returning to estrus or pregnancy rates. Weaning rate was significantly higher in pre-positive dams compared to negative ones (87.9 and 74.1%, respectively - P<0.05). The incidence of seroconversion was 16.5% (54/328). In seroconverted ewes, a high rate of return to estrus was observed (P≤0.05). The incidence of toxoplasmosis suggests production losses equivalent to 1.87% per year, in this system production. In addition, toxoplasmosis decreased the number of lambs per ewe. Therefore, to improve livestock performance, T. gondii infection should be monitored and controlled in the studied area.

RESUMO: Este trabalho tem como objetivo determinar a incidência da infecção pelo T. gondii em ovinos, durante o período gestacional e a extensão das perdas associadas. Amostras de sangue foram coletadas de 411 ovelhas Corriedale em dois rebanhos, em diferentes locais do Rio Grande do Sul, Brasil. Presença de T. gondii foi diagnosticada pela técnica de imunofluorescência indireta (RIFI). A soroprevalência no pré-acasalamento foi de 20,2%, sem diferença significativa entre os dois rebanhos estudados. A infecção por T. gondii foi influenciada pela idade (P ≤0,05). A soroprevalência no pré-acasalamento não influenciou a taxa de retorno ao estro e a taxa de prenhez. Taxa de desmame foi significativamente maior nas ovelhas positivas na pré-cobertura em comparação com as negativas (87,9 e 74,1%, respectivamente - P<0,05). A incidência de soroconversão foi de 16,5% (54/328). Em ovelhas que soroconverteram, uma alta taxa de retorno ao estro foi observada (P≤0,05). A incidência da toxoplasmose sugere perdas de produção equivalentes a 1,87% ao ano nestes sistemas. Além disso, a toxoplasmose diminuiu o número de cordeiros por ovelha. Portanto, para melhorar o desempenho produtivo nos rebanhos ovinos, a infecção por T. gondii deve ser monitorada e controlada na área estudada.

Verificação da transmissão vertical de Neospora spp. em equinos / Verification of vertical transmission of Neospora spp. in horses.

O gênero protozoário Neospora é reconhecido como causador de desordens reprodutivas e abortos em bovinos. Entre os equinos pouco se sabe sobre os efeitos da infecção por estes protozoários. Atualmente é admitido que os efeitos da infecção por Neospora hughesi em equinos possam ocorrer no sistema nervoso central e, os efeitos provocados pela infecção por Neospora caninum recaiam sobre o sistema reprodutor de éguas. O presente trabalho verificou a presença de imunoglobulinas da classe G no soro sanguíneo de uma população de éguas de cria e, em seus respectivos potros antes da ingestão do colostro. Para execução deste trabalho foi empregada técnica de imunofluorescência indireta (RIFI), utilizando como antígeno taquizoítos de Neospora caninum, a diluição inicial dos soros das éguas foi de 1:50 e a diluição do soro dos potros empregada foi de 1:16. Foram assistidos 78 partos e todos os potros tiveram seu soro sanguíneo coletado imediatamente após o nascimento. A pesquisa de anticorpos contra Neospora spp. apontou que 50 (64%) éguas e 32 (41%) potros foram positivos. Das 50 éguas que apresentaram anticorpos contra Neospora spp. 24 geraram potros positivos. Entre as 28 éguas que não reagiram, oito deram a luz a potros positivos. De posse dos resultados encontrados podemos concluir que ocorreu a transmissão vertical de Neospora spp. nos equinos pesquisados...

The genre protozoan Neospora is recognized as causing reproductive disorders and miscarriages in cattle. Among the horses little is known about the effects of infection by these protozoa. It is currently accepted that the effects of infection by Neospora hughesi in horses may occur in the central nervous system, and effects of Neospora caninum infection occur in the reproductive system of mares. The present study examined the presence of class immunoglobulin G in blood serum of a population of brood mares and their foals before colostrum ingestion. For this assignment was employed indirect immunofluorescence assay (IFA) using as antigen tachyzoites of Neospora caninum, the initial dilution employed in sera of the mares was 1:50 and dilution in the serum of foals was 1:16. Were assisted 78 deliveries and all foals had their blood serum collected immediately after birth. The presence of antibodies against Neospora spp. found in mares was 50 (64%) and 32 (41%) foals were positive. Of the 50 mares that had antibodies to Neospora spp. 24 generated positive foals. Among the 28 mares unreacted eight gave birth to foals positive. Having the results we can conclude that vertical transmission occurred Neospora spp. researched in horses...

Trichomonas Vaginalis Infection Among Women Attending in the Public Service in Rio Grande do Sul,Brazil: Frequency, Risk Factors and Clinical Signs / Infecção por Trichomonas Vaginalis em Mulheres a Tendidas pelo Serviço Público no Rio Grande do Sul, Brasil: Frequência, Fatores de Risco e Sinais Clínicos

Trichomoniasis is a cosmopolitan disease that can affect the female fertility, and is commonly underdiagnosed, both in private practices and in public health services, because of the low sensitivity of the wet mount exam used routinely. Objective: To understand the occurrence of T. vaginalis infection by means of in vitro culture in women receiving care in a public health unit, in the city of Pelotas, Rio Grande do Sul, Brazil, as well as to identify the possible risk factors associated with this infection. Methods: Cross­sectional study was carried out, which included 201 women undergoing interview and gynecological exam, with the collection of vaginal discharge in the Gynecology Ambulatory at the School of Medicine of Universidade Federal de Pelotas. The material collected was examined in the form of fresh smears and cultivated in Diamond's medium. Epidemiological data were obtained by means of patient interviews and clinical trials, from the medical records. The results were statistically analyzed through χ 2 and Fisher's exact tests, using version 9.0 of the Statistix program. Results: The occurrence of T. vaginalis infection was 7% (14/201). In the wet mount exam, used routinely for the diagnosis of this agent, only 42.85% of the infected women (6/14) were positive. It was noted that 21.4% of the infected women were asymptomatic, and 89.05% were not aware of the existence of the Trichomonas vaginalis infection. Factors independently associated with the infection were the smoking habit (odds ratio [OR] = 11.8), not having a stable sexual partner (OR = 6.36), presence of vaginal discharge with odor (OR = 5.65), and altered vaginal microbiota (OR = 5.31). Conclusion : T. vaginalis infection was present among the women studied, being underestimated because of the diagnostic technique, and because many of them were asymptomatic. The smoking habit, not having a stable sexual partner, having fetid discharge, and altered vaginal microbiota are the risk factors for infection.

A tricomoníase é cosmopolita, pode afetar a fertilidade feminina, e geralmente é subdiagnosticada, tanto em consultórios particulares, quanto em serviços públicos, devido à baixa sensibilidade do exame a fresco, usado rotineiramente. Objetivo: Conhecer a ocorrência de infecção por Trichomonas vaginalis , através de cultivo in vitro , em mulheres atendidas em unidade pública, na cidade de Pelotas, Rio Grande do Sul, além de identificar os possíveis fatores de risco associados a essa infecção. Métodos: Estudo de corte transversal que incluiu 201 mulheres, submetidas a entrevista e exame ginecológico, com coleta de conteúdo vaginal, no Ambulatório de Ginecologia da Faculdade de Medicina da Universidade Federal de Pelotas. O material coletado foi examinado a fresco e cultivado em meio de Diamond. Foram obtidos dados epidemiológicos através de entrevista, e clínicos, nos prontuários. Os resultados foram analisados estatisticamente através dos testes do χ 2 e exato de Fisher, utilizando o programa Statistix versão 9.0. Resultados: A ocorrência da infecção por T. vaginalis foi de 7% (14/201). No exame a fresco, usado como rotina para o diagnóstico desse agente, apenas 42,85% das infectadas (6/14) foram positivas. Constat ou­se que 21,4% das mulheres infectadas são assintomáticas, e que 89,05% desconhecia a existência da tricomoníase. Os fatores independent emente associados com a infecção foram o hábito de fumar (OR=11,8), não ter companheiro fixo (OR=6,36), apresentar corrimento vaginal fétido (OR=5,65) e microbiota vaginal alterada (OR=5,31). Conclusão: A infecção por T. vaginalis está presente entre as mulheres estudadas, e sendo subestimada, devido à técnica de diagnóstico e por muitas serem assintomáticas. O hábito de fumar, não ter companheiro fixo, ter corrimento fétido e microbiota vaginal alterada são fatores de risco para a infecção.