Érica Paes Barreto Xavier de Moraes

Experimental infection by Toxoplasma gondii using contaminated semen containing different doses of tachyzoites in sheep

Toxoplasmosis is a parasitic disease caused by Toxoplasma gondii that affects reproductive performance in small ruminants. Although the T. gondii life cycle is well understood since 1960s, several aspects related to its infection remain unclear. In the present study we hypothesized that sheep inseminated with T. gondii-contaminated semen would develop toxoplasmosis. In order to test that hypothesis, 41 sheep were experimentally infected with semen spiked with the organism. Females were divided in three groups (G1-G3): (a) females in G1 group were inseminated with semen containing 6.5 x 10(4) tachyzoites; (b) females in G2 group with semen containing 4 x 10(7) tachyzoites; and (c) females in G3 group with tachyzoite-free semen (control group). To confirm T. gondii infection via semen, serological tests were performed using indirect immunofluorescence reaction and the detection of parasite DNA in the blood stream using the nested PCR test. While in G1 group only 5/15 (33.3%) of the females presented seroconversion, all sheep in G2 15/15 (100%) seroconverted. The nested PCR test showed that 14/15 (93.3%) of the females in the G1 and 14/15 (93.3%) in the G2 group were positive for T. gondii while in the G3 group all samples were negative. In addition, ultra-sound test evidenced that in sheep presented embryonic reabsorption in animals from the infected groups. In conclusion, insemination using fresh semen experimentally contaminated with different infectant doses of T. gondii tachyzoites was able to infect sheep, leading to the possibility of toxoplasmosis transmission via semen.

Sexagem fetal em ovelhas Santa Inês por ultra-sonografia

The present study aimed to identify the sex and to determine the day of genital tubercle (GT) migration of ovine fetuses using real time ultrasonography. The sex was identified in Experiment (EI) taking into consideration the localization of GT and in Experiment II (EII) the presence of penis, prepuce and scrotal bag in male fetus and nipples, genital swelling and clitoris in female fetus. In EI, the females (n=17) were monitored with 12 hour intervals from the 35th to the 46th day of pregnancy, by transrectal via with linear transducer (6.0 and 8.0MHz). In EII, the females (n=30) with pregnancy period from 55 to 75 days were examined once only, using the same transducer and via used in EI. Among 17 females in EI, 11 (64.6%) fetuses were correctly sexed independent of single (7/11), twin (3/11) or triple (1/11) pregnancy In 6 (35.4%) pregnancies, 3 (17.7%) were twins, being impossible to sex one fetus of each pregnancy. In other 3 (17.7%) pregnancies the fetuses were correctly sexed, although the birth did not coincide with the quantification. In a male fetus of a single pregnancy, the migration of the GT began on day 37 of pregnancy and on the 46th day all the fetuses of the other pregnancies were correctly sexed. Among 30 females in EII, 16 (53.4%) pregnancies were single being sexed with accuracy of 100%. In other 14 (46.6%) remainder the pregnancies were twins, being impossible, in four cases, to be determined the sex of one of each twin. The incorrect diagnoses were fetuses sexed as females, however born as males. From the all born fetuses the total accuracy was 88.0% (EI) and 90.9% (EII), being not observed difference (P>0.05) between both experiments. The results allow to conclude that ultrasonography in real time is an efficient method to diagnose the fetal sex by visualization of GT, as well as by identification of penis, prepuce and scrotal bag in male fetus and nipples, genital swelling and clitoris in female fetus, since the scanning are performed from Day 50 of pregnancy

Detection of Toxoplasma gondii in the milk of naturally infected goats in the Northeast of Brazil

The aim of the present study was to detect the genomic DNA of Toxoplasma gondii in milk samples from naturally infected goats in the state of Pernambuco, (Brazil). In total, 248 blood serum samples were collected and processed from lactating goats and then submitted to a search for antibodies to T. gondii through the indirect immunofluorescence reaction. Samples with a score of 64 or more were considered positive. In total, 248 milk samples were collected and processed from the same group of goats in order to study the DNA of T. gondii using the polymerase chain reaction (PCR) technique. In the serum samples, 56/248 (22.58%) of the animals were positive, whereas the DNA of the parasite was detected in 15/248 (6.05%) of the milk samples. Five of these 15 samples were animals who were also positive in the serology. This study reports the first occurrence of the elimination of T. gondii from the milk of naturally infected goats in the north-east of Brazil. It is suggested that the consumption of in natura goat milk may constitute a potential risk to the health of milk consumers in this region.

Prevalence of Toxoplasma gondii in Slaughtered Pigs in the State of Pernambuco, Brazil

Abstract: The object of this study was to investigate Toxoplasma gondii antibodies and parasite DNA in pigs in the state of Pernambuco, Brazil. Blood samples were collected from 305 slaughtered pigs in 11 municipalities, and their sera were tested for T. gondii antibodies using the indirect fluorescent antibody test (IFAT, cutoff 1∶64); 38 (12.5%) samples were positive. Attempts were made to detect T. gondii DNA in the heart tissue of seropositive pigs using the B 1 gene and PCR; 21 (55.2%) of the 38 hearts were positive. This is the first detection of T. gondii DNA in tissues of serologically positive swine in the State of Pernambuco, Brazil.

Toxoplasma gondii diagnosis in ovine aborted fetuses and stillborns in the State of Pernambuco, Brazil.

The aim of this research was to study the contribution of Toxoplasma gondii to reproductive failure using nested PCR and histopathological examination of fetuses, stillborns and placentas. We examined 245 organs of fetuses and 28 placentas from 35 abortions and stillborns from naturally occurring miscarriages in sheep in the State of Pernambuco, Brazil. At necropsy, fragments of brain, cerebellum, medulla, lung, heart, spleen, liver and placenta were taken for nested PCR and histopathological tests. Pathological examination revealed macroscopic lesions, suggesting T. gondii infection in 5/35 (14.3%) of the placentas. The histopathological examination revealed no lesions characteristic of toxoplasmosis in the organs investigated. In the five placentas, lesions consistent with toxoplasmosis were observed as an inflammatory non-suppurative infiltrate, along with multiple necrosis and mineralization. Nested PCR showed three aborted fetuses and two stillborns (14.3%) to test positive for T. gondii, with DNA amplification in all organs and the placenta, especially the heart and the placenta, which are the tissues of choice. This study substantiates the theory that T. gondii is involved in miscarriages and stillbirths and in the placentas of naturally infected sheep in Brazil. Such findings have not previously been described in the national literature.

Experimental Vaginal Infection of Goats with Semen Contaminated with the “CPG” Strain of Toxoplasma gondii

Abstract:  The objective was to characterize the transmission of Toxoplasma gondii in goats experimentally infected vaginally with semen contaminated with the CPG strain (genotype III). Ten female goats were randomly allocated into 2 groups (G1 and G2), each with 5 animals, and inseminated during estrus. Goats in G1 were inseminated with semen containing 1 × 105 tachyzoites, whereas those in G2 (control) were inseminated with semen free from tachyzoites (insemination = day 0). In G1, seroconversion (indirect immunofluorescence reaction) and DNA (polymerase chain reaction) in the blood was present in 4/5 and 3/5, respectively, from the 7th day. In G2, all goats were negative in all tests. Embryonic reabsorption occurred in 4 of 5 goats from G1 between days 21 and 49. In conclusion, artificial vaginal insemination with semen containing tachyzoites of T. gondii–infected goats and is a potential transmission route of this parasite through semen.

Detecção de Toxoplasma gondii em órgãos do sistema reprodutivo de carneiros naturalmente infectados no Brasil

The aim of the study was to detect genomic DNA of Toxoplasma gondii in testicle and epididymis samples from rams sold in abattoirs in the state of Pernambuco, Northeast Brazil. Fifty (50) blood serum samples were collected, as well as 50 testicle and epididymis samples. Indirect Immunofluorescence (IIF) was used during screening of the rams. The Polymerase Chain Reaction (PCR) was used with animals that were positive in serology. Our results confirmed that 24% (12/50) of the rams were positive in IIF. Genomic DNA was detected in the epididymis at 8.3% (1/12) of the animals. The molecular identity of the amplified products was confirmed through sequencing. This paper reports the first occurrence of T. gondii DNA in the reproductive organs of naturally infected rams in Brazil.

Detection of Ureaplasma spp. in semen samples from sheep in Brazil

A study was conducted to verify the presence of mycoplasmas and ureaplasmas DNA in sheep semen samples from the State of Pernambuco. The PCR assay was conducted of according with standard protocols with generic primers. Mollicutes DNA was detected in 26.0% and Ureaplasma spp. in 12.0% of semen samples.

Occurrence of Toxoplasma gondii DNA in sheep naturally infected and slaughtered in abattoirs in Pernambuco, Brazil

The aim of the present study was to assess the occurrence of antibodies to Toxoplasma gondii and to detect genomic DNA of the parasite in the reproductive organs, fetuses and fetal membranes of sheep in slaughterhouses in the state of Pernambuco, Brazil. The Indirect Immunofluorescence technique (IFA) was used for screening. The Polymerase Chain Reaction (PCR) was used to detect DNA of T. gondii in the animals that were positive in the serology. In the serology, 13/50 samples were positive and genomic DNA of T. gondii was detected in one uterus, tube, ovary, placenta and fetus (heart, brain and umbilical cord) sample from a sheep that was positive in the serology. The present study provides evidence of the occurrence of T. gondii DNA in the organs of the reproductive system, placenta and fetus of a naturally infected sheep.

Ocorrência e fatores de riscos associados à infecção por Mycoplasma gallisepticum em galinhas na região Semiárida do estado de Pernambuco

The aim of the present study was to assess the occurrence of Mycoplasma gallisepticum (MG) infection and risk factors of this disease in three hundred serum samples from on 23 familiar agricultural properties in the semiarid region of the state of Pernambuco, Brazil. ELISA was used to study antibodies anti-Mycoplasma gallisepticum. The univariate analysis (chi-squared test or Fischers exact test) followed by multivariate analysis (logistic regression) were used to assess the risk factors with two variables: management and sanity of the poultry. It was detected a frequence of 53.33% (157/300) of the birds were positive for MG, with 100% foci. The risk factors confirmed by multivariate analysis, in the present study, were the presence of other poultry species on the property, including Numida meleagris (OR=2.22; p=0.005), parrots (OR=1.72; p=0.027), and of passerines (OR=1.88; p=0.007). These results showed that Mycoplasma gallisepticum infection is endemic among backyard poultry in the semiarid region of the state of Pernambuco. These birds could be a source of infection for other wild or domestic poultry. . This is the first report of the occurrence of avian mycoplasmosis in backyard poultry in the state of Pernambuco in northeastern Brazil. The risk factors identified should serve as a parameter for the health authorities to seek solutions related to controlling the disease.