Absence of Viable Toxoplasma gondii in Artisanal Raw-Milk Ewe Cheese Derived from Naturally Infected Animals

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.

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Physico-chemical analysis and detection of adulteration in raw milk collected from Goalas of different places of sadar upazila in Mymensingh district

Research in Agriculture Livestock and Fisheries ◽

10.3329/ralf.v4i2.33721 ◽

2017 ◽

Vol 4 (2) ◽

pp. 99-106

Author(s):

Atikur Rahman ◽

Md Rezwanul Habib ◽

Md Younus Ali ◽

Mohammad Ashiqul Islam ◽

Md Harun Ur Rashid

Keyword(s):

Research Work ◽

Raw Milk ◽

Physical Parameters ◽

Chemical Parameters ◽

Flowing Liquid ◽

Milk Samples ◽

Physical Chemical ◽

Physico Chemical ◽

White Colour ◽

Different Sources

This research work was carried out to assess the physical, chemical quality and detection of adulteration in raw milk collected from goals of five different places of Mymensingh sadar upazila (BAU Sheshmore, BAU KR market, train going vendor, sweetmeat shop and Dhudmohol) in Bangladesh. Results shows that milk from sweet meat shop had 100% yellowish white colour, normal (milky) flavor and free flowing liquid whereas other sources milk varies with their percentage in terms of physical parameters. Specific gravity of milk from various sources differed significantly (p<0.01) and all the chemical parameters of milk sample collected from different sources differed significantly (p<0.01). Significantly higher percentage of total solids (12.67±0.10) and fat content (4.36±0.07) was found in milk from goala of sweet meat shop than others. All of the adulteration tests showed negative result that means no adulterant materials was found in the collected milk samples. Considering the results, it could be inferred that there was an ample fluctuation present on qualities of milk samples collected from the goalas of different places of Mymensingh sadar regarding the standard of the parameters and the raw milk samples from sweetmeat shop were of good quality. The results suggested that milk purchase from reliable sources is very much important for consumers due to its quality.Res. Agric., Livest. Fish.4(2): 99-106, August 2017

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

10.21203/rs.3.rs-62998/v1 ◽

2020 ◽

Author(s):

Yong-Liang Wang ◽

QingFeng Meng

Keyword(s):

Toxoplasma Gondii ◽

Bovine Milk ◽

Raw Milk ◽

Human Infection ◽

Family Farming ◽

Blood Samples ◽

Milk Samples ◽

Fresh Milk ◽

Potential Source ◽

Low Prevalence

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.

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Prevalence estimation and genotypization of Toxoplasma gondii in goats

Biologia ◽

10.2478/s11756-010-0070-2 ◽

2010 ◽

Vol 65 (4) ◽

Cited By ~ 16

Author(s):

František Spišák ◽

Ľudmila Turčeková ◽

Katarína Reiterová ◽

Silvia Špilovská ◽

Pavol Dubinský

Keyword(s):

Toxoplasma Gondii ◽

Dna Polymorphism ◽

Age Groups ◽

Raw Milk ◽

Human Infection ◽

Molecular Methods ◽

Milk Samples ◽

Prevalence Estimation ◽

Genotype Ii

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.

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The Physicochemical Quality of Traditional Burduf Cheese

Bulletin of University of Agricultural Sciences and Veterinary Medicine Cluj-Napoca Food Science and Technology ◽

10.15835/buasvmcn-fst:9461 ◽

2013 ◽

Vol 70 (2) ◽

pp. 137

Author(s):

Carmen Pop ◽

Cristina Anamaria Semeniuc ◽

Sorin Apostu ◽

Ancuţa Mihaela Rotar

Keyword(s):

Quality Control ◽

Raw Milk ◽

Cow Milk ◽

Chemical Parameters ◽

Physicochemical Quality ◽

Milk Samples ◽

Sheep Milk ◽

Physico Chemical ◽

Different Types

The aim of this study is the assessmentof the quality control of raw milk and traditional burduf cheese obtained fromcow milk mixed with 10% sheep milk. Appreciation of the integrity and freshness assessmentof milk (cow and sheep) was tested by physico-chemical analysis.On theshelf-live were determined the physico-chemical parameters in cheese samples. Theantibiotics residues were tested of the milk samples with portable analyser,model Rosa Charm Reader. Theresults of physico-chemical determinations for the milk and cheese samples werewithin the maximum permissible by data legislation. Regardingthe content of antibiotics, the results were negative both for cow milk and forsheep milk. The sensorycharacteristics of burduf cheese are influenced by the different types of milk.

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Toxoplasma gondii and pre-treatment protocols for polymerase chain reaction analysis of milk samples: a field trial in sheep from Southern Italy

Italian Journal of Food Safety ◽

10.4081/ijfs.2017.6501 ◽

2017 ◽

Vol 6 (1) ◽

Cited By ~ 1

Author(s):

Alice Vismarra ◽

Elena Barilli ◽

Maura Miceli ◽

Carlo Mangia ◽

Cristina Bacci ◽

...

Keyword(s):

Toxoplasma Gondii ◽

Real Time ◽

Dna Extraction ◽

Real Time Pcr ◽

Southern Italy ◽

Raw Milk ◽

Type I ◽

Treatment Protocols ◽

Milk Samples ◽

Pre Treatment

Toxoplasmosis is a zoonotic disease caused by the protozoan Toxoplasma gondii. 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 T. gondii 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 T. gondii transmission through consumption of raw milk and its unpasteurized derivatives.

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Detection and Survival of Toxoplasma gondii in Milk and Cheese from Experimentally Infected Goats†

Journal of Food Protection ◽

10.4315/0362-028x.jfp-14-167 ◽

2014 ◽

Vol 77 (10) ◽

pp. 1747-1753 ◽

Cited By ~ 32

Author(s):

J. P. DUBEY ◽

S. K. VERMA ◽

L. R. FERREIRA ◽

S. OLIVEIRA ◽

A. B. CASSINELLI ◽

...

Keyword(s):

Risk Factor ◽

Toxoplasma Gondii ◽

Enzyme Treatment ◽

Mouse Bioassay ◽

Milk Samples ◽

Goat Cheese ◽

Goat’S Milk ◽

Unpasteurized Milk ◽

Fresh Cheese ◽

Goat's Milk

The consumption of unpasteurized goat cheese and goat's milk has been suggested as a risk factor for toxoplasmosis in humans. In the present study, detection and survival of Toxoplasma gondii in milk and cheese was studied by bioassay in mice (milk) and in cats (cheese). Eight goats were inoculated orally with 300 to 10,000 oocysts of T. gondii strain TgGoatUS26. Milk samples were collected daily up to 30 days postinoculation and bioassayed in mice and cats. For mouse bioassay, 50 ml of milk samples were centrifuged, and the sediment was inoculated subcutaneously into mice. Mice were tested for T. gondii infection by seroconversion and by the demonstration of parasites. By mouse bioassay, T. gondii was detected in milk from all eight goats. The T. gondii excretion in milk was intermittent. For cat bioassay, 400 ml (100 ml or more from each goat) of milk from four goats from 6 to 27 days postinoculation were pooled daily, and cheese was made using rennin. Ten grams of cheese was fed daily to four cats, and cat feces were examined for oocyst shedding. One cat fed cheese shed oocysts 7 to 11 days after consuming cheese. Attempts were made to detect T. gondii DNA in milk of four goats; T. gondii was detected by PCR more consistently, but there was no correlation between detection of viable T. gondii by bioassay in mice and T. gondii DNA by PCR. Results indicate that T. gondii can be excreted in goat's milk and can survive in fresh cheese made by cold-enzyme treatment. To prevent transmission to humans or animals, milk should not be consumed raw. Raw fresh goat cheese made by cold-enzyme treatment of unpasteurized milk also should not be consumed.

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Identification of Coxiella burnetii in Raw Milk of Livestock Animal in Iran

International Journal of Microbiology ◽

10.1155/2021/6632036 ◽

2021 ◽

Vol 2021 ◽

pp. 1-5

Author(s):

Ashraf Mohabati Mobarez ◽

Ehsan Mostafavi ◽

Mohammad Khalili ◽

Saber Esmaeili

Keyword(s):

Real Time ◽

Coxiella Burnetii ◽

Real Time Pcr ◽

Q Fever ◽

Goat Milk ◽

Raw Milk ◽

Molecular Evidence ◽

Milk Samples ◽

Sheep Milk ◽

Dairy Animals

Coxiella burnetii is the causative agent of Q fever in humans and animals. This study aimed to determine the frequency of C. burnetii in milk samples of dairy animals (goats, sheep, and cattle) in some selected regions in Iran, where there is no information about prevalence of C. burnetii. In this study, 162 individual milk samples were collected from 43 farms in three provinces (Tehran, Hamadan, and Mazandaran). Real-time PCR was used for the detection of IS1111a element of C. burnetii. In total, 23 of 162 samples (14.2%, 95% conﬁdence interval (CI): 9.65–20.2%) were positive for C. burnetii by real-time PCR. C. burnetii was detected in 10.17% (95% CI: 4.74–20.46) of goat milk samples. In sheep milk samples, 18.6% (95% CI: 9.74–32.62) were positive, and C. burnetii was detected in 15% (95% CI: 8.1–26.11) of cattle milk samples. Molecular evidence of the presence of C. burnetii was seen in milk samples of dairy animals in all the studied regions. These findings demonstrated that C. burnetii infection, especially in raw milk samples, deserves more attention from the health care system and veterinary organization in Iran.

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Molecular genetic methods of analysis to identify the species affiliation of the raw material composition in food products

Vestnik MGTU ◽

10.21443/1560-9278-2020-23-3-214-223 ◽

2020 ◽

Vol 23 (3) ◽

pp. 214-223

Author(s):

E. A. Yurova ◽

N. A. Zhizhin ◽

S. A. Filchakova

Keyword(s):

Dairy Products ◽

Diagnostic Method ◽

Molecular Genetic ◽

Food Products ◽

Raw Milk ◽

Biological Tissues ◽

Raw Material ◽

Dna Molecules ◽

Milk Samples ◽

Sheep Milk

Received in revised 08.08.2020 Methods based on the analysis of proteins and DNA molecules are more and more used to assess the composition of food products. Proteins research methods include immunological, electrophoretic and chromatographic ones. The analysis of DNA molecules is most often used to identify the species affiliation of food components. This is due to the stability of their structure compared to proteins, as well as their presence in most biological tissues. The results of studies evaluating methodological approaches for the application of the PCR diagnostic method to identify the composition of food products and the possibility of their use for monitoring dairy products have been shown. The objects of research were samples of cow, goat, sheep milk, as well as milk samples of different animal species mixed in various ratios. DNA was extracted from milk samples according to a unified technique for the separation of DNA molecules in milk and dairy products. The work also considers the possibility of using the PCR diagnostic method to identify the raw material origin of the product. To evaluate the measurement methods, artificially created samples of raw milk were used, which were cow, goat and sheep milk, a mix of three types of milk in different ratios. As a result of the research, the main method has been chosen as the real time PCR method, which has reliability, high sensitivity, sufficient rapidity, with the possibility of using it for dairy multicomponent products with a complex structural matrix, as well as products that have undergone deep technological processing.

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Quality of raw milk collected from Mymensingh town in Bangladesh

Bangladesh Journal of Animal Science ◽

10.3329/bjas.v45i2.29814 ◽

2016 ◽

Vol 45 (2) ◽

pp. 73-78 ◽

Cited By ~ 1

Author(s):

MS Uddin ◽

MR Habib ◽

MA Islam ◽

S Afrin ◽

MH Rashid

Keyword(s):

Specific Gravity ◽

Milk Fat ◽

Raw Milk ◽

Fat Content ◽

Chemical Parameters ◽

Total Solids ◽

Milk Samples ◽

Negative Results ◽

Different Sources

The present study was conducted to know the chemical qualities and adulteration of fresh raw milk collected from local markets (Bhangnamari bazaar, Sutiakhali bazaar, Vabokhali bazaar, Shombhuganj bazaar and Mymensingh sadar bazaar) of Mymensingh sadar, Bangladesh. Milk samples were analyzed for chemical parameters (contents of acidity, total solids, solids-not-fat, fat, protein and ash) and adulteration (presence of formalin, sugar or starch). Also, lactometer reading was recorded to monitor the specific gravity of the collected milk samples. Result showed that the specific gravity of milk from different sources differed non-significantly. The total solids, fat and protein contents of all the samples were in normal range except the milk fat content collected from Shombhuganj bazaar (32.66±4.04 g/Kg) and Mymensingh sadar bazaar (30.66±4.51 g/Kg) which had less (p<0.01) than the minimum legal standard of milk fat (35.00 g/Kg). The solids-not-fat content of the milk samples collected from Bhangnamari bazaar (72.93±8.78 g/Kg) and Vabokhali bazaar (79.33±6.81 g/Kg) were less than that from Mymensingh sadar bazaar (95.67±11.72 g/Kg). Developed acidity was detected in the samples from Bhangnamari bazaar (0.180±0.03), Sutiakhali bazaar (0.187±0.04%) and Mymensingh sadar bazaar (0.180±0.02%), while other samples were found to the fresh (0.150-0.157%). All of the adulteration tests showed negative results. The results suggested that though there were some fluctuations in quality among the bazaars regarding the standard of parameters, the raw milk samples were of acceptable quality.Bang. J. Anim. Sci. 2016. 45 (2): 73-78

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Detection of antibacterial residues in milk by HPLC-DAD and microbial inhibitor tests

Czech Journal of Food Sciences ◽

10.17221/262/2018-cjfs ◽

2020 ◽

Vol 38 (No. 1) ◽

pp. 63-71

Author(s):

Sofia Christoforidou ◽

Eftychia Karageorgou ◽

Maria Ioannidou ◽

Evdoxios Psomas ◽

Martha Maggira ◽

...

Keyword(s):

Raw Milk ◽

Hplc Method ◽

Detection Sensitivity ◽

Detection Capability ◽

Milk Samples ◽

Sheep Milk ◽

Microbial Inhibition ◽

Maximum Residue Levels ◽

Array Detection ◽

Residue Levels

The objective of this study was to assess the detection sensitivity of four commercial microbial inhibition assays (MIAs) in comparison with the results by High Pressure Liquid Chromatography with Diode Array Detection (HPLC-DAD) for residues of three tetracyclines, trimethoprim and sulfadiazine in fortified raw milk samples. MIAs showed fairly good results at detecting sulfadiazine residues, whereas concerning tetracyclines and trimethoprim residues, most of the kits showed insufficient results. HPLC analysis revealed high recoveries of examined compounds with detection limits below their respective maximum residue levels (MRLs). The HPLC method was validated determining linearity, accuracy, precision, detection capability and decision limit. Subsequently, both methods were used to monitor several cow and sheep milk samples for the presence of antibacterial agents. Results obtained showed that MIAs could be successfully used for the analysis of different milk types. However, milk spoilage which was observed in some cases could decrease the sensitivity of the kits, a fact that does not seem to influence the effectiveness of HPLC-DAD method.

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