scholarly journals Short Reports Low Prevalence of Toxoplasma Gondii DNA in the Fresh Milk of Cattle in China

2020 ◽  
Author(s):  
Yong-Liang Wang ◽  
QingFeng Meng

Abstract Background: It is generally recognized that there is a risk for humans to acquire toxoplasmosis through consumption of raw milk from infected animals. Although several studies have been conducted to detect the seroprevalence of T. gondii in cattle in China, no study has been conducted to detect T. gondii DNA in milk of cattle in China. Thus, the present study was firstly conducted to explore the prevalence of T. gondii DNA in fresh milk of cattle in China. Results: A total of 2092 blood samples and fresh bovine milk were collected from six provinces between January 2018 and June 2019, respectively, and examined by ELISA and semi-nested PCR, respectively. In total, 123/2092 (5.88%) of the blood samples and 22/2092 (1.05%) of the milk samples were positive for T. gondii, respectively. Fifteen of these 22 positive milk samples (68.18%) were animals who also recorded serologically positive. Moreover, cattle and milk from family farming have significantly higher T. gondii prevalence than those from commercial farm. Conclusions: To our knowledge, this is the first report of T. gondii DNA was found in the fresh milk of cattle in China, suggesting that the consumption of raw milk from seropositive cattle could be a potential source of human infection.

2020 ◽  
Vol 8 (1) ◽  
pp. 143 ◽  
Author(s):  
David Ranucci ◽  
Elena Battisti ◽  
Fabrizia Veronesi ◽  
Manuela Diaferia ◽  
Giulia Morganti ◽  
...  

The presence of viable Toxoplasma gondii was investigated in artisanal cheeses made from milk of naturally infected ewes. Ewe milk was analyzed beforehand for the presence and vitality of T. gondii by loop-mediated isothermal amplification (LAMP) and reverse-transcriptase PCR (RT-PCR), respectively. Cheeses were prepared from raw milk following a traditional cheesemaking process. The cheese obtained from T. gondii-positive milk was analyzed by LAMP to detect Toxoplasma DNA-positive samples. RT-PCR was then carried out to assess the viability of the parasites in T. gondii-positive milk samples and fresh cheese, after 5 and 15 days of ripening. Physical-chemical parameters of cheeses were also investigated. All cheese samples derived from T. gondii-positive milk were positive according to LAMP, at both 5 and 15 days of ripening, while none of the samples were positive according to RT-PCR. Thus, while the presence of the parasite was demonstrated by the detection of specific DNA, the absence of detectable T. gondii RNA supports the hypothesis that changes in the chemical and physical characteristics occurring during the cheesemaking process and ripening period, could be sufficient to inactivate viable T. gondii in milk, minimizing the risk of human infection through consumption of raw sheep milk cheese.


Biologia ◽  
2010 ◽  
Vol 65 (4) ◽  
Author(s):  
František Spišák ◽  
Ľudmila Turčeková ◽  
Katarína Reiterová ◽  
Silvia Špilovská ◽  
Pavol Dubinský

AbstractIn this study we aimed to determine seroprevalence of antibodies against Toxoplasma gondii and parasite DNA presence in the milk of goats from a farm in eastern Slovakia. Anti-Toxoplasma antibodies were detected in 43 goat sera out of 87 examined (49.43%). The highest prevalence was recorded in the goats aged more than 72 months (76.19%; OR = 4.62; 95% CI = 1.51-14.14) and the lowest one in animals aged from 12 to 36 months (17.65%; OR = 0.09; 95% CI = 0.03-0.27). Statistically significant correlation (P < 0.0001) was found between the prevalence of antibodies against T. gondi and animal age in comparing age groups - goats up to 36 months of age and above 37 months of age. The presence of T. gondii DNA was confirmed in 32.56% of milk samples using molecular methods. Based on the DNA polymorphism at the SAG2 locus of T. gondii we identified the goats as being infected with genotype II of T. gondii. Presence of DNA in milk refers to the risk of human infection through consuming raw milk.


2021 ◽  
Vol 9 (5) ◽  
pp. 1097
Author(s):  
Labrini V. Athanasiou ◽  
Vasileios G. Papatsiros ◽  
Victoria M. Spanou ◽  
Eleni G. Katsogiannou ◽  
Anna Dedousi

Neospora caninum and Toxoplasma gondii affect both humans and animals worldwide. To investigate their seroprevalence and differences in seropositivity between pigs vaccinated and unvaccinated against porcine circovirus 2 (PCV2), as well as differences in muscle enzyme activity between seropositive and seronegative pigs, blood samples were collected from 380 sows. Antibodies against T. gondii and N. caninum were detected by an indirect immunofluorescence antibody (IFA) assay, while the activities of creatine kinase (CK) and aspartate aminotransferase (AST) were biochemically assessed. Out of the 364 sows finally included in the study, 4.4%, 3.5%, and 0.5% were seropositive to T. gondii, N. caninum, or both. A significantly higher percentage of seropositivity against T. gondii and/or N. caninum in PCV2 unvaccinated pigs compared with vaccinated pigs was observed. Increased serum activities of CK and AST were detected in 71.43% and 100% of only against T. gondii (T+) and 63.64% and 90.91% of only against N. caninum (N+) seropositive sows, respectively, and were significantly higher compared to seronegative animals. T. gondii and N. caninum seropositivity, especially in presumed immunocompromised pigs, and the evidence of muscle damage highlight their importance as a zoonotic pathogen and animal model of human infection, respectively.


2017 ◽  
Vol 6 (1) ◽  
Author(s):  
Alice Vismarra ◽  
Elena Barilli ◽  
Maura Miceli ◽  
Carlo Mangia ◽  
Cristina Bacci ◽  
...  

Toxoplasmosis is a zoonotic disease caused by the protozoan <em>Toxoplasma gondii</em>. Ingestion of raw milk has been suggested as a risk for transmission to humans. Here the authors evaluated pre-treatment protocols for DNA extraction on <em>T. gondii</em> tachyzoite-spiked sheep milk with the aim of identifying the method that resulted in the most rapid and reliable PCR positivity. This protocol was then used to analyze milk samples form sheep from three different farms in southern Italy, including Real Time PCR for DNA quantification and PCR-RFLP for genotyping. The pre-treatment protocol using EDTA and Tris-HCl to remove casein gave the best results in the least amount of time compared to the others on spiked milk samples. One sample of 21 collected from sheep farms was positive on one-step PCR, Real Time PCR and resulted in a Type I genotype at one locus (SAG3). Milk usually contains a low number of tachyzoites and this could be a limiting factor for molecular identification. Our preliminary data has evaluated a rapid, cost-effective and sensitive protocol to treat milk before DNA extraction. The results of the present study also confirm the possibility of <em>T. gondii</em> transmission through consumption of raw milk and its unpasteurized derivatives.


2018 ◽  
Vol 38 (8) ◽  
pp. 1577-1583 ◽  
Author(s):  
José C. Ribeiro Júnior ◽  
Pedro I. Teider Junior ◽  
André L.M. Oliveira ◽  
Edson A. Rios ◽  
Ronaldo Tamanini ◽  
...  

ABSTRACT: Pseudomonas, the main genus of gram-negative microorganisms isolated from milk, is psychrotrophic, biofilm-forming, and thermo-resistant deteriorating enzyme producers. The aim of this study was to quantify Pseudomonas spp. in goat’s and cow’s milk produced in the Paraná state, Brazil, to evaluate the deteriorating activity of the isolates at mesophilic and psychrotrophic conditions and to identify, at the species level, the isolates with alkaline metalloprotease (aprX gene) production potential. Microbiological, biochemical and molecular methods were used for isolating, confirming and identifying of isolates. The mean counts were 1.6 (±6.3)x104 and 0.89(±3)x102 CFU/mL for goat and bovine milk samples, respectively, immediately after milking. Of the Pseudomonas colonies isolated from goat milk (n=60), 91.7% showed proteolytic potential when incubated at 35°C/48 h and 80% at 7°C/10 days, and lipolytic potential was observed in 95% of the isolates incubated in mesophilic and 78.3% at refrigeration conditions. From the isolates of bovine milk (n=20), 35% showed proteolytic activity only when incubated at 35°C/48 h, and lipolytic potential was observed in 25% of the isolates incubated at 7°C/10d and 35°C/48h. It was observed that 83.3% and 25% of the isolates genetically confirmed as Pseudomonas spp. of goat and bovine milk showed the potential for alkaline metalloprotease production, with the species P. azotoformans, P. koreensis, P. gessardii, P. monteilii and P. lurida being the most frequent in goat milk and P. aeruginosa the only species identified in cow milk.


1976 ◽  
Vol 43 (2) ◽  
pp. 251-258 ◽  
Author(s):  
B. J. Kitchen

SummaryAssay procedures were developed for a number of enzymes in milk which apparently originate from leucocytes. The enzymes studied were acid phosphatase, N-acetyl-β-D-glucosaminidase, β-glucuronidase, arylsulphatase, α-mannosidase, and catalase. Quarter-milk samples were analysed for enzyme activity and results compared with the electronic cell count and the Wisconsin Mastitis Test. All enzymes measured except acid phosphatase and α-mannosidase showed good correlation with the electronic cell count. Of the other 4 enzymes tested, β-glucuronidase and arylsulphatase were unsuitable as diagnostic aids owing to the lengthy incubation periods required in their assay procedures. The assay of catalase, which involved the measurement of the initial rate of release of O2 using an O2 analyser apparatus, was rapid, sensitive and reasonably reliable, if fresh milk samples were used. The assay procedure for N-acetyl-β-D-glucosaminidase was considered to be the most reliable, simple and rapid enzymic method for estimating the number of somatic cells in milk.


Parasite ◽  
2018 ◽  
Vol 25 ◽  
pp. 21 ◽  
Author(s):  
Mike Heddergott ◽  
Natalia Osten-Sacken ◽  
Peter Steinbach ◽  
Alain C. Frantz

Despite increasing consumption of mouflon (Ovis orientalis musimon) meat in Germany, there is currently no surveillance of Toxoplasma gondii infection in populations of these animals and generally little knowledge about the prevalence of this protozoan in German wild ungulates. Between 2011 and 2015, we collected 138 blood samples from a free-living mouflon population in central German and tested sera for the presence of T. gondii antibodies using a modified agglutination test (MAT, cut-off 1:20). Antibodies were detected in 31 of the 138 samples (22.46%). There was a significant difference in seroprevalence between the different age classes, with antibodies to T. gondii more frequent in adults. In contrast, there was no significant difference in seroprevalence depending on sex and year of sample collection. Game meat is frequently consumed as raw or undercooked meat and may therefore represent a potential source of human infection with T. gondii.


2009 ◽  
Vol 92 (3) ◽  
pp. 959-974 ◽  
Author(s):  
Mohamed Abouzied ◽  
Michael Sarzynski ◽  
Aaron Walsh ◽  
Heather Wood ◽  
Mark Mozola

Abstract Avalidation study designed tomeet the requirements of the AOAC Research Institute and the U.S. Food and Drug Administration (FDA), Center for Veterinary Medicine, was conducted for a receptor-based, immunochromatographic method (BetaStar US) for detection of beta-lactam antibiotic residues in raw, commingled bovine milk. The assay was found to detect amoxicillin, ampicillin, cephapirin, cloxacillin, and penicillin G at levels below the FDA tolerance/safe levels but above the maximum sensitivity thresholds established by the National Conference on Interstate Milk Shipments. Results of the Part I (internal) and Part II (independent laboratory) dose-response studies using spiked samples were in very close agreement for all five drugs tested, with differences between the Part I and Part II 90/95 sensitivity values ranging from 0 to 1 ppb. The test was able to detect all five drugs at the approximate 90/95 sensitivity levels when present as incurred residues in milk collected from cows that had been treated with the specific drug. Asixth drug, ceftiofur, was found to be undetectable at levels of 500 ppb (as total ceftiofur metabolites from incurred residues in milk samples). The selectivity of the assay was 100, because no false-positive results were obtained in tests of &gt;1000 control milk samples. The assay was found to be applicable to the testing of frozen raw milk samples. Results of ruggedness experiments established the operating parameter tolerances for the BetaStar US assay. Results of cross-reactivity testing established that the assay detects certain other beta-lactam drugs (dicloxacillin and ticarcillin), but it does not cross-react with any of 30 drugs belonging to other classes. Abnormally high bacterial or somatic cell counts in rawmilk produced no interference with the ability of the test to detect beta-lactams at tolerance/safe levels.


2019 ◽  
Vol 40 (2) ◽  
pp. 723
Author(s):  
Amanda Bezerra Bertolini ◽  
Marcelo Augusto Orsi Dutra ◽  
Lívia Maísa Guiraldi ◽  
Wesley José dos Santos ◽  
Maria Izabel Merino de Medeiros ◽  
...  

Milk and its derivatives are good substrates for the proliferation of pathogenic and quality-deteriorating microorganisms, demanding rigorous care with milking, processing, and storage. Among the various bacteria that can grow in raw refrigerated milk, Yersinia enterocolitica, is an invasive enteropathogen of humans. This bacterium can cause a number of intestinal and extraintestinal clinical symptoms, ranging from mild gastroenteritis to mesenteric lymphadenitis, similar to appendicitis. To evaluate the prevalence of pathogenic Yersinia enterocolitica in raw milk from bulk milk tanks located in the State of São Paulo, 102 bovine milk samples (one per dairy farm) were evaluated by microbiological analyses, followed by biochemical tests PCR and genetic sequencing. Microbiological testing did not isolate Y. enterocolitica. However, PCR analysis revealed six samples that were positive for Y. enterocolitica (5.9%), confirmed by genetic sequencing. Only the inv gene was detected, which is present in virulent and avirulent Y. enterocolitica strains. There was great difficulty in microbiological isolation due to the difficulty of competitiveness of Y. enterocolitica in a very rich microbiota of raw milk. Although virulence genes known to be present in potentially pathogenic strains of Y. enterocolitica have not been identified, the presence of this pathogen in milk from expansion tanks, identified through PCR and confirmed by genetic sequencing, suggests that Y. enterocolitica may be a risk to public health, especially if milk and its derivatives are consumed without heat treatment.


2020 ◽  
Vol 3 (2) ◽  
pp. 1-9
Author(s):  
IEM El Zubeir ◽  

This study was designed to investigate the effect of using the recommended FAO Lactoperoxidase Enzyme System (LPS) on improving the keeping quality and increasing the shelf life of raw milk from camels at different stages of lactations. Fresh milk samples were obtained after morning milking from Camel Research Center of Khartoum University.


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