Goat Paratuberculosis: Experimental Model for the Evaluation of Mycobacterium Persistence in Raw Milk Cheese

Mycobacterium avium subsp. paratuberculosis (MAP) is the causative agent of chronic proliferative enteritis found in ruminants, known as paratuberculosis (PTB). The spread of PTB is increasing in countries with advanced animal husbandry practices, leading to significant economic losses. Moreover, a supposed zoonotic role of MAP in Crohn’s disease (CD) in humans has been discussed by the scientific community; however, although the association between MAP and CD has generally been accepted, it is still up for debate if MAP is the main cause of CD, a contributing factor, or merely a commensal organism for the development of CD. The aim of this study was to assess the survival of MAP during the entire production process of a traditional Italian goat’s raw milk fresh cheese, the “Robiola di Roccaverano”, assessing the survival rate and persistence of MAP in the final product. A mix of MAP field isolates from goats of the Roccaverano area and a reference ATCC strain were used to carry out milk in experimental inoculation. Samples of milk, curd and cheese were taken in two consecutive batches of production. Microbiological challenge tests, evaluated by f57-qPCR, showed a significant decrease in MAP charge during the cheesemaking process for both batches, suggesting the productive process has an impact on MAP survival.

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Virulence and Immunity Orchestrated by the Global Gene RegulatorsigLin Mycobacterium avium subsp. paratuberculosis

Infection and Immunity ◽

10.1128/iai.00001-14 ◽

2014 ◽

Vol 82 (7) ◽

pp. 3066-3075 ◽

Cited By ~ 9

Author(s):

Pallab Ghosh ◽

Howard Steinberg ◽

Adel M. Talaat

Keyword(s):

Mycobacterium Avium ◽

Critical Role ◽

Biological Significance ◽

Sigma Factors ◽

Economic Losses ◽

Bacterial Survival ◽

Content Type ◽

Vaccine Potential ◽

Mycobacterium Avium Subsp Paratuberculosis

ABSTRACTMycobacterium aviumsubsp.paratuberculosiscauses Johne's disease in ruminants, a chronic enteric disease responsible for severe economic losses in the dairy industry. Global gene regulators, including sigma factors are important in regulating mycobacterial virulence. However, the biological significance of such regulators inM. aviumsubsp.paratuberculosisrremains elusive. To better decipher the role of sigma factors inM. aviumsubsp.paratuberculosispathogenesis, we targeted a key sigma factor gene,sigL, activated in mycobacterium-infected macrophages. We interrogated anM. aviumsubsp.paratuberculosisΔsigLmutant against a selected list of stressors that mimic the host microenvironments. Our data showed thatsigLwas important in maintaining bacterial survival under such stress conditions. Survival levels further reflected the inability of the ΔsigLmutant to persist inside the macrophage microenvironments. Additionally, mouse infection studies suggested a substantial role forsigLinM. aviumsubsp.paratuberculosisvirulence, as indicated by the significant attenuation of the ΔsigL-deficient mutant compared to the parental strain. More importantly, when thesigLmutant was tested for its vaccine potential, protective immunity was generated in a vaccine/challenge model of murine paratuberculosis. Overall, our study highlights critical role ofsigLin the pathogenesis and immunity ofM. aviumsubsp.paratuberculosisinfection, a potential role that could be shared by similar proteins in other intracellular pathogens.

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698. Effect of the heat treatment of milk on the yield, quality and certain properties of standardized separated milk cheese

Journal of Dairy Research ◽

10.1017/s0022029900009067 ◽

1958 ◽

Vol 25 (1) ◽

pp. 85-91 ◽

Cited By ~ 9

Author(s):

G. M. El Sadek ◽

L. Abd El Motteleb

Keyword(s):

Sodium Chloride ◽

Total Nitrogen ◽

Raw Milk ◽

The Body ◽

Pasteurized Milk ◽

Milk Whey ◽

Rennet Coagulation ◽

Fresh Cheese ◽

Yield Quality ◽

Raw Milk Cheese

1. Four batches of soft cheese were made using rennet coagulation from both raw and pasteurized separated milk standardized to contain 1·5% fat and 7% added sodium chloride.2. The yield of pasteurized milk cheese was about 3·33% higher than that of raw milk cheese.3. Pasteurization of the milk improved the flavour and the texture of the fresh cheese, but had no marked effects on the body and general appearance.4. The shelf keeping property after 4–5 days under the conditions of these experiments was better for the cheese made from raw milk. Reasons for this apparent anomaly are discussed.5. Chemical analyses showed a greater moisture retention by the pasteurized milk cheese, leading to a lower content of fat and total nitrogen but increased sodium chloride in solution. Some retention of heat coagulable protein in the pasteurized milk cheese was suggested by a higher total nitrogen in dry-matter figure and by a lower nitrogen content of the pasteurized milk whey.6. Pasteurization of the milk greatly reduced the total bacterial population of the fresh cheese and also the acidity of the whey produced.

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Serological survey of paratuberculosis in dairy cattle in Garfagnana district (Tuscany)

Journal of the Hellenic Veterinary Medical Society ◽

10.12681/jhvms.16067 ◽

2018 ◽

Vol 68 (4) ◽

pp. 641 ◽

Cited By ~ 1

Author(s):

A. GALIERO ◽

F. FRATINI ◽

P. BRAVI ◽

B. TURCHI ◽

E. CASANOVI ◽

...

Keyword(s):

Bovine Milk ◽

Raw Milk ◽

Economic Losses ◽

Pcr Analysis ◽

Human Consumption ◽

Serum Samples ◽

Confirmation Test ◽

Mycobacterium Avium Subsp Paratuberculosis ◽

Serological Data

Mycobacterium avium subsp. paratuberculosis (Map) is the agent of a chronic, progressive granulomatous enteritis in ruminants known as Johne’s disease or paratuberculosis. Nowadays, the interest regarding this pathogen is increasing not only because Map causes economic losses, but it has also been suggested as a potential risk factor for the development of human diseases such as Crohn disease, autoimmune diseases like type-1 diabetes, sarcoidosis, multiple sclerosis and Hashimoto’s thyroiditis. The aim of our study was to determine the presence of paratuberculosis in Garfagnana (Tuscany, Italy) where raw milk is not submitted to pasteurization, but it is directly sold to consumers or it is destined for cheese making without the employment of heat treatment. The survey was conducted in Garfagnana district where there are 17 herds which produce and market bovine milk for direct human consumption. Serum samples (n=162) were obtained form 16 herds and were analyzed performing ELISA ID screen® Paratuberculosis Indirect screening test (ID.VET, Montpellier, France) and positive samples were tested with ELISA ID screen® Paratuberculosis Indirect confirmation test (ID.VET, Montpellier, France), according to the instructions provided by the manufacturer. The analysis performed by ELISA revealed that the true seroprevalence was 29.1% at the herd level and 4.6% at the animal level. In our opinion, although ELISA is a very useful tool to screen herds for paratuberculosis, more studies should be carried out in our territory combining serological data with cultural and PCR analysis.

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Raw milk cheese as a potential infection source of pathogenic and toxigenic food born pathogens

Food Science and Technology ◽

10.1590/fst.37919 ◽

2020 ◽

Author(s):

Marita Vedovelli CARDOZO ◽

Natalia NESPOLO ◽

Tammy Chioda DELFINO ◽

Camila Chioda de ALMEIDA ◽

Lucas José Luduverio PIZAURO ◽

...

Keyword(s):

Raw Milk ◽

Infection Source ◽

Raw Milk Cheese

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Estimation of Aflatoxin M1 Levels in Some Dairy Products Manufactured from Raw Milk Experimentally Inoculated with Toxin

The Iraqi Journal of Veterinary Medicine ◽

10.30539/iraqijvm.v43i1.471 ◽

2019 ◽

Vol 43 (1) ◽

pp. 50-58

Author(s):

H. S. Alnaemi

Keyword(s):

Dairy Products ◽

Raw Milk ◽

Aflatoxin M1 ◽

Consumer Health ◽

Goat’S Milk ◽

White Cheese ◽

Elisa Technique ◽

Permissible Levels ◽

Goat's Milk

Fate of AflatoxinM1 in soft white cheese and its by-product (whey) and in yogurt locally made from raw sheep's and goat's milk experimentally inoculated with 0.05 and 0.5 µg/l AflatoxinM1 were investigated using ELISA technique. Results reported that AflatoxinM1 was concentrated in cheese at levels significantly higher than that recorded in the raw milk that used for its processing, with a significant decrease in AflatoxinM1 levels in its by-product (whey) comparable to the raw milk used in manufacturing at both inoculated levels. Yogurt produced from raw sheep's milk at second inoculated level exerted AflatoxinM1concentration significantly lower than that present in the milk. Significant differences in AflatoxinM1distribution in cheese and whey produced from sheep's milk comparable to their counterparts produced from goat's milk were recorded. Finally, results revealed the efficacious role of the various dairy manufacturing processes in AflatoxinM1 distribution and the necessity to issue of local legislations concerning the maximum permissible limits for AflatoxinM1 in milk in order to stay within the universal permissible levels for AflatoxinM1 in dairy products to provide greater protection for consumer health.

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Impact of Raw Materials and Processing Techniques on The Microbiological Quality of Egyptian Domiati Cheese

International Journal of Veterinary Science ◽

10.37422/ijvs/20.060 ◽

2020 ◽

Keyword(s):

Raw Materials ◽

Microbiological Quality ◽

Raw Milk ◽

Plate Count ◽

E Coli ◽

Fresh Cheese ◽

Processing Techniques ◽

The Impact ◽

Total Yeast

Domiati cheese is the most popular brand of cheese ripened in brine in the Middle East in terms of consumed quantities. This study was performed to investigate the impact of the microbiological quality of the used raw materials, the applied traditional processing techniques and ripening period on the quality and safety of the produced cheese. Three hundred random composite samples were collected from three factories at Fayoum Governorate, Egypt. Collected samples represent twenty-five each of: raw milk, table salt, calf rennet, microbial rennet, water, environmental air, whey, fresh cheese, ripened cheese & swabs from: worker hands; cheese molds and utensils; tanks. All samples were examined microbiologically for Standard Plate Count (SPC), coliforms count, Staphylococcus aureus (S. aureus) count, total yeast & mould count, presence of E. coli, Salmonellae and Listeria monocytogenes (L. monocytogenes). The mean value of SPC, coliforms, S. aureus and total yeast & mould counts ranged from (79×102 CFU/m3 for air to 13×108 CFU/g for fresh cheese), (7×102 MPN/ cm2 for tank swabs to 80×106 MPN/ml for raw milk), (9×102 CFU/g for salt to 69×106 CFU/g for fresh cheese) and (2×102 CFU/cm2 for hand swabs to 60×104 CFU/g for fresh cheese), respectively. Whereas, E. coli, Salmonella and L. monocytogenes failed to be detected in all examined samples. There were significant differences in all determined microbiological parameters (p ≤0.05) between fresh and ripened cheese which may be attributed to different adverse conditions such as water activity, pH, salt content and temperature carried out to improve the quality of the product.

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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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Evaluating the Authenticity of the Raw-Milk Cheese Fontina (PDO) with Respect to Similar Cheeses

Foods ◽

10.3390/foods10020350 ◽

2021 ◽

Vol 10 (2) ◽

pp. 350

Author(s):

Luisa Pellegrino ◽

Johannes A. Hogenboom ◽

Veronica Rosi ◽

Paolo D’Incecco

Keyword(s):

Amino Acids ◽

Free Amino Acids ◽

Free Amino ◽

Raw Milk ◽

Manufacturing Processes ◽

Food Authenticity ◽

Promising Tool ◽

Production Area ◽

The Mean ◽

Raw Milk Cheese

The implementation of quality assurance schemes for the assessment of PDO food authenticity is an issue involving manufacturers, traders, retailers and consumers. In this respect, reliable analytical methods are needed to integrate paper-trailing information. The feasibility of distinguishing the Italian Fontina PDO cheese from the generic Fontal cheese was preliminarily evaluated on a set of commercial samples by measuring selected parameters (pH, alkaline phosphatase activity, content of copper, volatiles, extent of proteolysis) related to the different manufacturing processes. The relative profile of free amino acids proved to be a promising tool. A new set of 41 samples of Fontina PDO cheese was collected at representative dairies within the recognized production area and analyzed for free amino acids. A chemometric model of Fontina PDO cheese was built based on the mean content and standard deviation of 15 free amino acids. On this basis, all of the PDO samples were correctly identified, whereas all of the Fontal cheeses were recognized as different cheeses.

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Analysis of the Role of Bradysiaimpatiens (Diptera: Sciaridae) as a Vector Transmitting Peanut Stunt Virus on the Model Plant Nicotiana benthamiana

Cells ◽

10.3390/cells10061546 ◽

2021 ◽

Vol 10 (6) ◽

pp. 1546

Author(s):

Marta Budziszewska ◽

Patryk Frąckowiak ◽

Aleksandra Obrępalska-Stęplowska

Keyword(s):

Nicotiana Benthamiana ◽

Molecular Mechanisms ◽

Virus Transmission ◽

Pathogenic Fungi ◽

Economic Losses ◽

Fungus Gnats ◽

Droplet Pcr ◽

Viral Coat ◽

Peanut Stunt Virus

Bradysia species, commonly known as fungus gnats, are ubiquitous in greenhouses, nurseries of horticultural plants, and commercial mushroom houses, causing significant economic losses. Moreover, the insects from the Bradysia genus have a well-documented role in plant pathogenic fungi transmission. Here, a study on the potential of Bradysia impatiens to acquire and transmit the peanut stunt virus (PSV) from plant to plant was undertaken. Four-day-old larvae of B. impatiens were exposed to PSV-P strain by feeding on virus-infected leaves of Nicotiana benthamiana and then transferred to healthy plants in laboratory conditions. Using the reverse transcription-polymerase chain reaction (RT-PCR), real-time PCR (RT-qPCR), and digital droplet PCR (RT-ddPCR), the PSV RNAs in the larva, pupa, and imago of B. impatiens were detected and quantified. The presence of PSV genomic RNA strands as well as viral coat protein in N. benthamiana, on which the viruliferous larvae were feeding, was also confirmed at the molecular level, even though the characteristic symptoms of PSV infection were not observed. The results have shown that larvae of B. impatiens could acquire the virus and transmit it to healthy plants. Moreover, it has been proven that PSV might persist in the insect body transstadially. Although the molecular mechanisms of virion acquisition and retention during insect development need further studies, this is the first report on B. impatiens playing a potential role in plant virus transmission.

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