Lice (Haematopinus tuberculatus) in water buffalo farms from central Italy

The aim of the present study was to obtain information about the presence and distribution of the suckling louse Haematopinus tuberculatus in water buffalo farms in central Italy. The survey was carried out on 127 farms (epidemiological units), selected using a grid approach within a Geographical Information System, followed by proportional allocation. In each farm 6 buffaloes were examined in order to detect the louse presence. Parasitological examinations were performed on each buffalo at predilection sites. A total of 762 water buffaloes were examined. H. tuberculatus was found in the 11.0% (14/127) of the farms and in the 4.5% (34/762) of the animals. The presence H. tuberculatus should be routinely considered because it is a cause of serious health, production and economic damages in intensive breeding buffaloes

Droplet Digital PCR (ddPCR) Analysis for the Detection and Quantification of Cow DNA in Buffalo Mozzarella Cheese

Buffalo mozzarella cheese is one of the most appreciated traditional Italian products and it is certified as a Protected Designation of Origin (PDO) product under the European Commission Regulation No. 1151/2012. It is obtained exclusively from buffalo milk. If made from cow milk, or a mixture of buffalo and cow milk, buffalo mozzarella cheese does not qualify as a PDO product. In order to maximize their profits, some producers market buffalo mozzarella that also contains cow milk as a PDO product, thus defrauding consumers. New methods for revealing this fraud are therefore needed. One such method is the droplet digital Polymerase Chain Reaction (ddPCR). Thanks to its high precision and sensitivity, the ddPCR could prove an efficacious means for detecting the presence of cow milk in buffalo mozzarella cheese that is marketed as a PDO product. ddPCR has proved able to detect the DNA of cow and/or buffalo milk in 33 buffalo mozzarella cheeses labelled as PDO products, and experimental evidence could support its application in routine analyses

Trichinella britovi in wild boar meat from Italy, 2015–2021: A citizen science approach to surveillance

As a result of the increase of game meat intended for human consumption through Europe, a plethora of food-borne diseases, including trichinellosis, may occur in consumers, posing a relevant public health threat.Thus, this study aims to a citizen science approach to monitor the occurrence of Trichinella spp. in wild boar meat intended for human consumption, evaluating the risk of infection for consumers.Following the European Regulation 2015/1375 (laying down specific rules on official controls for Trichinella in meat), from 2015 to 2021, hunters (n = 478) were involved to collect diaphragm pillar samples of wild boars from mainland southern Italy, which were tested for Trichinella spp. L1 larvae via HCl-pepsin digestion and Multiplex PCR.Overall, 139,160 animals were collected (average of 19,880 per year), being 14 (i.e., 0.01%) tested positive to Trichinella britovi by the combined biochemical and molecular approach. An average larval burden of 28.4 L1 per gram of meat was found (minimum 3.2 - maximum 132.6). A statistically significant difference was found in the prevalence according to hunting seasons (p < 0.01, with higher values in 2016 and 2021) and regions of the study area (p < 0.01). No statistically significant decrease in the prevalence of T. britovi throughout the study period was found (p = 0.51), except in Apulia region (p < 0.01).These findings revealed a stable prevalence of T. britovi in wild boar meat intended for human consumption, suggesting a risk of infection for consumers, especially hunters and local markets users. Citizen science surveillance models could be promoted to improve trichinellosis control and prevention in a One Health perspective

The microbiota of water buffalo milk during mastitis

<div><p>The aim of this study was to define the microbiota of water buffalo milk during sub-clinical and clinical mastitis, as compared to healthy status, by using high-throughput sequencing of the 16S rRNA gene. A total of 137 quarter samples were included in the experimental design: 27 samples derived from healthy, culture negative quarters, with a Somatic Cell Count (SCC) of less than 200,000 cells/ml; 27 samples from quarters with clinical mastitis; 83 samples were collected from quarters with subclinical mastitis, with a SCC number greater of 200,000 cells/ml and/or culture positive for udder pathogens, without clinical signs of mastitis. Bacterial DNA was purified and the 16S rRNA genes were individually amplified and sequenced. Significant differences were found in milk samples from healthy quarters and those with sub-clinical and clinical mastitis. The microbiota diversity of milk from healthy quarters was richer as compared to samples with sub-clinical mastitis, whose microbiota diversity was in turn richer as compared to those from clinical mastitis. The core microbiota of water buffalo milk, defined as the asset of microorganisms shared by all healthy milk samples, includes 15 genera, namely <i>Micrococcus</i>, <i>Propionibacterium</i>, <i>5-7N15</i>, <i>Solibacillus</i>, <i>Staphylococcus</i>, <i>Aerococcus</i>, <i>Facklamia</i>, <i>Trichococcus</i>, <i>Turicibacter</i>, <i>02d06</i>, <i>SMB53</i>, <i>Clostridium</i>, <i>Acinetobacter</i>, <i>Psychrobacter</i> and <i>Pseudomonas</i>. Only two genera (<i>Acinetobacter</i> and <i>Pseudomonas</i>) were present in all the samples from sub-clinical mastitis, and no genus was shared across all in clinical mastitis milk samples. The presence of mastitis was found to be related to the change in the relative abundance of genera, such as <i>Psychrobacter</i>, whose relative abundance decreased from 16.26% in the milk samples from healthy quarters to 3.2% in clinical mastitis. Other genera, such as <i>SMB53</i> and <i>Solibacillus</i>, were decreased as well. Discriminant analysis presents the evidence that the microbial community of healthy and clinical mastitis could be discriminated on the background of their microbiota profiles.</p></div

Beta diversity analysis.

<p>Unweighted Unifrac analysis including H (Healthy) and CM (Clinical mastitis) samples. Adonis: R = 0.17 p = 0.001 ANOSIM: R = 0.37 <i>p</i> = 0.001. Panel A: results including H, SM and CM quarters. Panel B: results including only H and CM. o = CM; + = SM; Δ = H</p

Water buffalo milk microbiota composition at the genus level for the 16S rRNA gene after classification of clinically healthy samples in SCC classes.

<p>Microbiota composition at the genus level for the 16S rRNA gene: Class 1, with a SCC < 100,000, Class 2, with a SCC between 100,000 and 500,000, Class 3, with a SCC between 500,000 and 1,000,000 and Class 4, with a SCC > 100,000,000.</p

Alpha diversity analysis.

<p>Rarefaction curves of samples with regards to quarter patho-physiological status (CM: clinical mastitis; H: healthy; SM: sub-clinical mastitis), as defined by the Shannon index. Statistical difference is present between H and CM groups (<i>p</i> = 0.03).</p

Relative abundance of microbiota taxa at phylum level.

<p>Relative abundance of microbiota taxa at phylum level.</p

Water buffalo milk taxonomic profile at genus level.

<p>The microbial relative abundance at genus level between: H = Healthy samples; SM = Sub-Clinical mastitis samples; CM = Clinical mastitis samples; * indicates statistical significance (<i>p</i> ≤ 0.05).</p