Molecular characterization of Toxoplasma gondii and Sarcocystis spp. in raw kibbeh and other meat samples commercialized in Botucatu, Southeastern Brazil

Abstract Toxoplasmosis occurs worldwide causing economic losses to the animal production and problems to the public health. The study aimed to detect Toxoplasma gondii and Sarcocystis spp.in 141 meat products from commercial meat cuts of pork, beef, and kibbeh sold in commercial markets from Botucatu, SP, Brazil. Samples were bioassayed in mice to isolate the parasite, and the parasite DNA detected by PCR targeting the 529 base pairs repeat element region (PCR-529-bp). All samples resulted negative on bioassay, whereas PCR positive for 9 (6,38%), distributed as 5/48 beef, 3/49 pork, and 1/44 kibbeh. PCR-positive were investigated for the the parasite genotype using multiplex-, nested-, and RFLP-PCR for 11 markers (SAG1, 5’-3’SAG2, alt.SAG2, SAG3, B-TUB, GRA6, L358, c22-8, c29-6, PK1, Apico). Complete genotype was determined on just one PCR-positive sample that matched MAS, TgCkBr89 and TgCkBr147 isolates already identified. In addition, nested- and RFLP-PCR targeting 18S rRNA was run for all PCR-positive samples and, the products, sequenced and aligned to the GenBank at NCBI website. Four samples showed 100% homology with T. gondii (GenBank #L37415.1), three with Sarcocystis hominis (GenBank #AF006471.1), two Sarcocystis cruzi (GenBank #AF176934.1), and one Sarcocystis hirsuta (GenBank #AF006469.1), indicating the circulation of T. gondii and Sarcocystis spp.

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Relación entre las características de los transportes, con las características nutricionales de la carne porcina destinada a consumo humano en el Valle de Aburrá, 2017

Revista de la Facultad de Medicina Veterinaria y de Zootecnia ◽

10.15446/rfmvz.v64n3.68692 ◽

2017 ◽

Vol 64 (3) ◽

Author(s):

Natalia Uribe ◽

Catalina María Arango ◽

Juan Fernando Naranjo ◽

Ángela Maria Segura ◽

Santiago Henao

Keyword(s):

Water Supply ◽

Animal Welfare ◽

Meat Products ◽

Economic Losses ◽

Human Consumption ◽

Chi Square ◽

Nutritional Parameters ◽

Probabilistic Sampling ◽

Qualitative Variables ◽

Meat Samples

Pork meat is considered a source of high nutritional value due to its high protein content, however, transport is a critical link to generate economic losses by producing alterations in animal welfare, which have an impact on nutritional parameters, decreasing the capacity of water retention, and generating protein losses. The objective of this study was to relate the characteristics of transport, with the nutritional characteristics of pork for human consumption in the Valle de Aburrá in 2017. Three slaughterhouses of Valle de Aburrá were visited with probabilistic sampling, stratified by plant and equal affixation, obtaining information from 338 animals. The nutritional parameters of the meat samples and sociodemographic variables, infrastructure, animal welfare and driving practices in the transporters were analyzed. Chi square tests were performed for dichotomous qualitative variables, logistic regression for qualitative polytomous variables and U Mann - Whitney for quantitative variables. An association was found between several of the parameters investigated with statistically significant p values (p = 0.000), where, having no permanent water supply for the animals increases the possibility of presenting nutritionally inadequate meats 46.55 times (IC 18.08 - 120.07). It concludes that factors such as lack of water supply to pigs, poor condition of the floors and separators, lack of training in transporters, lack of supervision of animals, lack of mechanical technical certification and average speed of 80 Km/Hr are associated with the generation of nutritionally inadequate meat products.

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Molecular detection of Toxoplasma gondii in aborted fetuses of goats in Chattogram, Bangladesh

Veterinary World ◽

10.14202/vetworld.2021.2386-2391 ◽

2021 ◽

pp. 2386-2391

Author(s):

Tanjila Hasan ◽

Abdul Mannan ◽

Delower Hossain ◽

Azizunnesa Rekha ◽

Md. Monir Hossan ◽

...

Keyword(s):

Toxoplasma Gondii ◽

Congenital Toxoplasmosis ◽

Positive Sample ◽

Reproductive Failure ◽

Control Measures ◽

Farm Animals ◽

Economic Losses ◽

Livestock Industry ◽

Nested Polymerase Chain Reaction ◽

Congenital Diseases

Background and Aim: Toxoplasma gondii is a protozoan parasite that is responsible for the major cause of congenital diseases, abortion, and stillbirth in humans and farm animals. Primary infection in pregnant goats due to T. gondii leads to abortion and significant economic losses in the livestock industry. Moreover, very few studies have been performed for the identification of T. gondii from aborted fetuses of goats. The study was conducted for the molecular identification of Toxoplasma gondii from aborted fetuses of goats in Chattogram, Bangladesh. Materials and Methods: Twenty aborted fetuses of goats were collected from 52 farms in the study area. A nested polymerase chain reaction (PCR) assay targeting the B1 gene was performed, and a positive sample yield of 197 bp amplified DNA products consistent with T. gondii. Results: The overall prevalence of toxoplasmosis in the aborted fetus of goats was 35.0%. Heart muscle, liver, brain, and placenta showed positive PCR results. The risk factors related to the does age, presence of cats in farms, and aborted fetus age were found to be statistically significant (p<0.05). Our results showed that T. gondii is a major possible causal factor for abortion and reproductive failure in goats. The high prevalence of T. gondii infection in aborted fetuses of goats revealed that T. gondii could be imperative in causing reproductive failure in goats. Conclusion: Active or congenital toxoplasmosis was shown by the presence of T. gondii in fetal tissues, which is a matter of concern as this parasite has zoonotic significance and causes economic hazards to the livestock industry by causing various reproductive problems. Therefore, proper control measures and strategies are needed to reduce the rate of abortion in goats, ultimately saving the livestock industry.

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Sarcocystidae in wild birds of southeastern Brazil

Revista Brasileira de Parasitologia Veterinária ◽

10.1590/s1984-296120201094 ◽

2021 ◽

Vol 30 (1) ◽

Author(s):

Wagner Martins Fontes do Rêgo ◽

Júlia Gatti Ladeia Costa ◽

Ramon Castro de Araujo Baraviera ◽

Lorena Velozo Pinto ◽

Gabriella de Lima Bessa ◽

...

Keyword(s):

Toxoplasma Gondii ◽

Ribosomal Dna ◽

Nested Pcr ◽

Minas Gerais ◽

The State ◽

Wild Birds ◽

Southeastern Brazil ◽

Sarcocystis Spp ◽

Rescue Center

Abstract This study aimed to identify members of the Sarcocystidae family in naturally infected wild birds at a rescue center in the state of Minas Gerais, southeastern Brazil. The heart and brain of 44 wild birds were evaluated by bioassay in mice to detect T. gondii, and extracted DNA was used for nested PCR of the 18S ribosomal DNA gene to detect members of the Sarcocystidae family. The positive samples were sequenced, assembled, edited and compared with sequences deposited in GenBank. Toxoplasma gondii was isolated from six (13.6%) out of 44 birds. Toxoplasma gondii DNA was identified in 10/44 (22.7%) of the birds. The amplified sequences exhibited 100% similarity with the DNA of the ME49 strain of T. gondii. Sarcocystis DNA (99% similarity) was identified in 5/44 (11.4%) of the birds. T. gondii and Sarcocystis spp. are common in wild birds in Minas Gerais, Brazil.

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Molecular detection of Apicomplexa protozoa in tissues from Alouatta guariba clamitans

Pesquisa Veterinária Brasileira ◽

10.1590/1678-5150-pvb-6717 ◽

2021 ◽

Vol 41 ◽

Author(s):

Aline Ludwig ◽

Laurete Murer ◽

Helton F. dos Santos ◽

Adriana Ludwig ◽

Luis Antonio Sangioni ◽

...

Keyword(s):

Toxoplasma Gondii ◽

Neospora Caninum ◽

Positive Sample ◽

Primate Species ◽

Howler Monkey ◽

Rrna Gene ◽

Tissue Samples ◽

Genetic Sequencing ◽

Sarcocystis Spp ◽

Alouatta Guariba Clamitans

ABSTRACT: The brown howler monkey (Alouatta guariba clamitans) is a primate species widely distributed in South America. Infections by protozoa are common in primates. However, studies on protozoa in primates in Brazil are scarce, so the goal of this study was to investigate DNA from the apicomplexan protozoa Neospora caninum, Sarcocystis spp. and Toxoplasma gondii in tissues of A. guariba clamitans. DNA extraction was performed on tissue samples from the heart, brain, liver, spleen, lung and intestine of six A. guariba clamitans from Santa Maria, Central Region of Rio Grande do Sul, Brazil. Conventional PCR was performed using 18S rRNA gene general primers for Apicomplexa and also specific primers to amplify Neosporaspp. and Toxoplasma gondii DNA. All animals were positive in the 18S PCR and the genetic sequencing confirmed the presence of Sarcocystis spp. DNA in the tissues of four animals belonging to at least two species (S. neurona and S. gigantea) and T. gondii DNA in the other two animals. One positive sample for T. gondii was genotypically characterized as atypical by the restriction fragment length polymorphism technique. N. caninum DNA was not detected in the tested samples. The presence of Apicomplexa protozoan DNA in the tissues of the six animals tested in this study highlights the importance of howler monkeys as maintainers of these pathogens in nature.

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Comparison of an Isothermal Amplification and Bioluminescence Detection of DNA Method and ISO 6579:2002 for the Detection of Salmonella enterica Serovars in Retail Meat Samples

Journal of Food Protection ◽

10.4315/0362-028x.jfp-12-313 ◽

2013 ◽

Vol 76 (4) ◽

pp. 657-661 ◽

Cited By ~ 2

Author(s):

SILVIA BONARDI ◽

IRENE ALPIGIANI ◽

CRISTINA BACCI ◽

FRANCO BRINDANI ◽

STEFANO PONGOLINI

Keyword(s):

Salmonella Enterica ◽

Meat Products ◽

Storage Period ◽

Positive Sample ◽

Isothermal Amplification ◽

Most Probable Number ◽

Probable Number ◽

Retail Meat ◽

Meat Samples ◽

Positive Results

The aim of the study was the comparative evaluation of an isothermal amplification and bioluminescence detection of DNA (IMBD) method and method ISO 6579:2002 for detection of Salmonella in retail meat products of unknown contamination status. A total of 200 meat samples were tested: 116 minced meat and meat preparations to be eaten cooked (52 chicken, 48 pork, and 16 beef samples) and 84 fresh meat samples (68 poultry and 16 pork). With one or both methods, 21 samples (10.5%) were positive for Salmonella enterica. Fifteen samples were positive with both methods (71.4% of all positive samples), two more samples (9.5%) were positive with the IMBD method only, and four samples (19.1%) were positive with the ISO method only. One ISO-positive sample was inhibited with the IMBD method. For the IMBD method, relative accuracy was 97.0% (95% confidence interval [CI], 93.6 to 98.9%), relative sensitivity was 78.9% (95%CI, 54.4 to 93.9%), and relative specificity was 98.9% (95% CI, 96.1 to 99.7%). Time to negative results was shorter with the IMBD method (20 to 24 h). Also, positive results were available in 20 to 24 h but should be confirmed using other methods (presumptive-positive results). Rapidity of response of the IMBD method gave us the opportunity to test the presumptive-positive samples by the most-probable-number (MPN) method, which was not performed for samples that were positive only with the ISO method because of likely microbial changes during the long storage period (5 to 7 days) at refrigeration temperature. Salmonella MPN values in naturally contaminated meat were low, at <0.3 to 2.1 MPN/g.

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Shiga toxin-producing Escherichia coli in meat and vegetable products in Emilia Romagna Region, years 2012-2013

Italian Journal of Food Safety ◽

10.4081/ijfs.2015.4511 ◽

2015 ◽

Vol 4 (1) ◽

Cited By ~ 4

Author(s):

Lia Bardasi ◽

Roberta Taddei ◽

Lucia Nocera ◽

Matteo Ricchi ◽

Giuseppe Merialdi

Keyword(s):

Escherichia Coli ◽

Shiga Toxin ◽

Virulence Genes ◽

Meat Products ◽

Gene Isolation ◽

E Coli ◽

Vegetable Products ◽

Emilia Romagna Region ◽

Meat Samples ◽

Pcr Targeting

In 2012-2013 Emilia-Romagna Region introduced a monitoring plan for Shiga toxin-producing Escherichia coli (STEC) in foodstuff. Six hundred eighty-nine meat samples and 273 fruit and vegetable products were analyzed according to ISOTS13136. Pre-enriched samples were tested by multiplex real time PCR targeting the virulence genes eae, stx1 and stx2. Stx2 positive samples were investigated for the presence of serogroup O104 associated gene. O103, O111, O145, O157, O26 associated genes were tested on samples positive for stx in association with eae gene. Isolation of E. coli strains was attempted from samples positive for serogroup-associated genes. Thirtyfour meat products (4.9%) resulted positive for stx1 and/or stx2 genes and 46 (6.7%) for stx1 and/or stx2 genes in association with eae gene. Forty-five (6.5%) samples resulted positive at least at one serogroup. Serogroup O103, O104, O111, O145, O157 and O26 genes were detected respectively in 1.3, 0.3, 0.1, 3.9, 2.9 and 2.5% samples; 0.6% samples resulted positive for STEC isolation (2 E. coli O103 and 2 E. coli O157). It is worth noting that STEC virulence genes were detected at high frequency (19%) in fresh pork meat sausages. Four (1.5%) vegetable samples were positive for stx1 and/or stx2 genes and 1 (0.4%) for stx1 and/or stx2 genes in association with eae gene; none resulted positive for the tested serogroups. Only a low number of samples positive by molecular methods were confirmed by cultural isolation. It is therefore of the uttermost importance for appropriate risk management, to be fully aware of the meaning of the analytical result.

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Monitoring of the Listeria spp. identification from the poultry products in the Dnipropetrovsk region

Scientific Messenger of LNU of Veterinary Medicine and Biotechnology ◽

10.32718/nvlvet9318 ◽

2019 ◽

Vol 21 (93) ◽

pp. 103-108

Author(s):

N. M. Zazharska ◽

I. V. Borovuk

Keyword(s):

Meat Products ◽

Food Contamination ◽

Biochemical Properties ◽

Food Plants ◽

Economic Losses ◽

Microbiological Analysis ◽

Normative Documents ◽

Differential Identification ◽

Food Taking ◽

Meat Samples

Due to high mortality, listeriosis is one of the most common causes of death from illnesses associated with food, taking the second place after salmonellosis. Listeriosis, as a rule, arises as a result of consumption of contaminated products, including meat products, cheese, ready-to-eat foods. L. monocytogenes belongs to the third group of pathogenicity. Contamination by L. monocytogenes in processing of products is a constant problem in food plants. Food contamination Listeria leads to a withdrawal of products that produces economic losses. Analysis of the dynamic detection and of the differential identification of Listeria spp. in the meat products of poultry processing enterprises in Dnipropetrovsk region was conducted. The research was carried out by Dnipropetrovsk regional state laboratory of the state service of Ukraine for food safety and consumer protection. The results of bacteriological researches of meat samples which poultry plants gave for microbiological analysis during period 2008–2018 were used for monitoring. Microbiological research was carried out in accordance with valid international normative documents. The fluorescence analyzer Mini Vidas, France, the CAMP test were used for analysis. The biochemical properties of isolated microorganisms were established using BioMerieux API tests, France. Analyzing the number of researches and identification of microorganisms in the Dnipropetrovsk region for the period of 11 years, 3001 positive results out of 8172 analyzed samples were found (36.7%). Herewith, part of positive samples goes up from 8.5% in 2008 to 77.9% in 2018. L. ivanovii was isolated in 1523 samples (18.6%), L. inocua – 833 (10.2%), L. monocytogenes – 493 (6%), L. seeliger – 97 (1.2%), L. grayi – 36 (0.4%), L. welshimeri in 19 samples of meat products (0.2%) out of the 8172 microbiological studies conducted over 11 years. Of the six types of identified Listeria, more than half were L. ivanovii, which is twice as high as cases with L. incocua and thrice compared to L. monocytogenes.

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Molecular detection of cattle Sarcocystis spp. in North-West Italy highlights their association with bovine eosinophilic myositis

Parasites & Vectors ◽

10.1186/s13071-021-04722-5 ◽

2021 ◽

Vol 14 (1) ◽

Author(s):

Selene Rubiola ◽

Tiziana Civera ◽

Felice Panebianco ◽

Davide Vercellino ◽

Francesco Chiesa

Keyword(s):

18S Rdna ◽

Inflammatory Myopathy ◽

Molecular Techniques ◽

Diaphragm Muscle ◽

Economic Losses ◽

Intermediate Hosts ◽

Slaughter Cattle ◽

Significant Difference ◽

Pcr Targeting

Abstract Background Cattle are intermediate hosts of six Sarcocystis species, among which Sarcocystis hominis and Sarcocystis heydorni can infect humans through the consumption of raw or undercooked meat. In addition to the zoonotic potential, there is increasing interest in these protozoa because of the evidence supporting the role of Sarcocystis spp. in the occurrence of bovine eosinophilic myositis (BEM), a specific inflammatory myopathy which leads to carcass condemnation and considerable economic losses. Actually, all the prevalence studies carried out on cattle in Italy have been based on either morphological or 18S rDNA-based molecular techniques, most likely leading to misidentification of closely related species. Therefore, there is a strong need for new data on the prevalence of the different Sarcocystis spp. in cattle in Italy and their association with bovine eosinophilic myositis. Methods To reach our aim, individual striated muscle samples from BEM condemned carcasses (N = 54) and diaphragm muscle samples from randomly sampled carcasses (N = 59) were obtained from Northwest Italy slaughterhouses. Genomic DNA was extracted and analyzed by multiplex-PCR targeting 18S rDNA and cox1 genes. PCR products amplified using the genus-specific primer set in absence of the specific fragment for S. hirsuta, S. cruzi, S. hominis or S. bovifelis were sequenced to achieve species identification. Results Sarcocystis DNA was detected in 67.8% of the samples from slaughter cattle and in 90.7% of the samples from BEM condemned carcasses. S. cruzi was identified as the most prevalent species in slaughter cattle (61%), followed by S. bovifelis (10.2%), S. hominis (8.5%) and S. hirsuta (1.7%). Notably, among the different Sarcocystis spp. detected, the presence of S. bovifelis and S. hominis was significantly higher in samples isolated from BEM condemned carcasses (46.3% and 40.7% respectively), while there was no statistically significant difference between the presence of S. cruzi or S. hirsuta in BEM condemned carcasses (42.6% and 1.8%, respectively) and randomly sampled carcasses. Furthermore, DNA sequence analysis revealed the presence of a putative new species in two carcasses. Conclusions Our study contributes to updating the data on the prevalence of the different Sarcocystis spp. in cattle in Italy, highlighting the presence of three Sarcocystis spp., S. cruzi, S. hominis and S. bovifelis, in BEM lesions and allowing us to speculate on the possible role of S. hominis and S. bovifelis as the major sarcosporidian species involved in bovine eosinophilic myositis. Graphic Abstract

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Seroprevalence of Toxoplasma gondii and associated risk factors in domestic pigs raised from Cuba

Parasitology Research ◽

10.1007/s00436-021-07245-1 ◽

2021 ◽

Author(s):

Julio César Castillo-Cuenca ◽

Álvaro Martínez-Moreno ◽

José Manuel Diaz-Cao ◽

Angel Entrena-García ◽

Jorge Fraga ◽

...

Keyword(s):

Risk Factors ◽

Toxoplasma Gondii ◽

Meat Products ◽

Control Measures ◽

Health Concern ◽

Enzyme Linked Immunosorbent Assay ◽

Serum Samples ◽

Cross Sectional ◽

Weaning Pigs ◽

Associated Risk Factors

AbstractA cross-sectional study was carried out to determine the seroprevalence of Toxoplasma gondii and associated risk factors in pigs in the largest pork-producing region in Cuba. Serum samples from 420 pigs, including 210 sows and 210 post-weaning pigs, were tested for antibodies against T. gondii using a commercial indirect enzyme-linked immunosorbent assay. Anti-T. gondii antibodies were detected in 56 animals (13.3%, 95% CI: 10.1–16.6). A generalized estimating equations model revealed that the risk factors associated with higher seropositivity in pigs were altitude (higher in farm’s location < 250 m above sea level (masl) versus ≥ 250 masl) and age (higher in sows compared to post-weaning pigs). The results indicated that this protozoan parasite is widely distributed on pig farms in the study area, which is a public health concern since the consumption of raw or undercooked pork meat products containing tissue cysts is considered one of the main routes of T. gondii transmission worldwide. Control measures should be implemented to reduce the risk of exposure to T. gondii in pigs in Cuba.

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Prevalence and Genetic Characterisation of Human Sapovirus from Children with Diarrhoea in the Rural Areas of Vhembe District, South Africa, 2017–2020

Viruses ◽

10.3390/v13030393 ◽

2021 ◽

Vol 13 (3) ◽

pp. 393

Author(s):

Mpho Magwalivha ◽

Jean-Pierre Kabue Ngandu ◽

Afsatou Ndama Traore ◽

Natasha Potgieter

Keyword(s):

South Africa ◽

Rural Communities ◽

Rural Areas ◽

Rna Extraction ◽

Diarrhoeal Disease ◽

Positive Sample ◽

Prevalent Strain ◽

Stool Samples ◽

Vhembe District ◽

Pcr Targeting

Diarrhoeal disease is considered an important cause of morbidity and mortality in developing areas, and a large contributor to the burden of disease in children younger than five years of age. This study investigated the prevalence and genogroups of human sapovirus (SV) in children ≤5 years of age in rural communities of Vhembe district, South Africa. Between 2017 and 2020, a total of 284 stool samples were collected from children suffering with diarrhoea (n = 228) and from children without diarrhoea (n = 56). RNA extraction using Boom extraction method, and screening for SV using real-time PCR were done in the lab. Positive samples were subjected to conventional RT-PCR targeting the capsid fragment. Positive sample isolates were genotyped using Sanger sequencing. Overall SV were detected in 14.1% (40/284) of the stool samples (16.7% (38/228) of diarrhoeal and 3.6% (2/56) of non-diarrhoeal samples). Significant correlation between SV positive cases and water sources was noted. Genogroup-I was identified as the most prevalent strain comprising 81.3% (13/16), followed by SV-GII 12.5% (2/16) and SV-GIV 6.2% (1/16). This study provides valuable data on prevalence of SV amongst outpatients in rural and underdeveloped communities, and highlights the necessity for further monitoring of SV circulating strains as potential emerging strains.

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