Mastitis is a highly prevalent condition that has a great impact on milk production and animal welfare, and often requires substantial management efforts. For this reason, it is generally considered an important threat to the dairy industry. Many microbial, host, and environmental factors can protect against, predispose to, or influence the development of mastitis. The objective of this work was to characterize the milk microbiota of Manchega ewes, and to compare samples from animals with and without a history of mastitis. We analyzed milk samples from 36 ewes belonging to 2 different farms (18 ewes from each farm) using culture-dependent and culture-independent techniques. We also analyzed several immune compounds to investigate associations of mastitis with 3 main variables: farm; history of mastitis or no mastitis; and parity number. Both culture-dependent and culture-independent techniques showed that ewe milk harbored a site-specific complex microbiota and microbiome. Staphylococcus epidermidis was the main species driving the difference between farm A (where it was the dominant species) and B (where it was not). In contrast, samples from farm B were characterized by the presence of a wide spectrum of other coagulase-negative staphylococci. Some of these species have already been associated with subclinical intramammary infections in ruminants. Of the 10 immune compounds assayed in this study, 3 were related to a history of mastitis [IL-8, IFN-γ, and IFN-gamma-induced protein 10 (IP-10)]. Increases in IL-8 concentrations in milk seemed to be a feature of subclinical mastitis in sheep, and in this study, this immune factor was detected only in samples from ewes with some episodes of mastitis and from the group with the highest somatic cell count. We also observed a positive correlation between the samples with the highest somatic cell count and IFN-γ and IP-10 levels. Our results suggest that these 3 compounds could be used as biomarkers for the negative selection of mastitis-prone animals, particularly when somatic cell count is very high.

Yeasts are always present in any type of cheese, as well as in the factories where it is produced. However, the role of the yeast community in the cheese making process, as well as the routes of contamination used by yeast species to contaminate milk from the dairy farm environment, are not well known. The objectives of this study were to broaden the knowledge of the dominant yeast community in Manchega sheep’s milk and to assess the contamination routes of the yeast species depending on the farm practices. Milk, teat surface (collected from ten ewes per farm), feed, and air (collected in milking parlours and livestock housing) samples were collected from 12 typical farms in Castilla-La Mancha, Spain with differences in farming practices, and the yeast species were identified using DNA sequencing methods. To evaluate whether certain farming practices have an effect on the distribution of species of yeast in the milk samples, a mixed model was used. The results showed that most of the dominant yeast species (mainly belonging to the genus Candida) found in milk were also found in the other samples, indicating a microbial transfer from the farm environment to the milk. Furthermore, the statistical model showed that factors influencing yeast counts in milk were the presence of yeasts in the milking parlour, the use of silage, and the frequency of acid treatment for cleaning the milking machines. In conclusion, milk contamination from the yeast species present in the dairy farm environment is related to certain farming practices such as the use of silage and the daily use of acid in the cleaning of the milking machines, which favours the presence of desirable microbiota in milk.

Over the past few years, there has been a great development of the sheep dairy sector in Spain and a wide variability in the production outcomes of farms, although most of the farms have similar environmental and economic constraints. Milk quality evaluation is important not only for cheese production, but also for other dairy products. Therefore, the aim was to evaluate the effect of some factors on bulk-tank milk characteristics of Manchega sheep, with special attention to casein and urea contents and colour indices. Factors of variation such as season, annual milk yield of ewe (AMYE), the enrolment (or not) in a breeding programme (BPM) and the feeding strategy of farms have been considered. The study included 77 farms distributed along La Mancha region, in Spain. Data about production practices and feeding strategy were also collected from these farms. A total of 308 analytical determinations of milk characteristics including fat, protein, total solids, casein, lactose, protein/total solids, casein/fat, pH, urea content, colour indices and somatic cell count were analysed by automated methods. The results of the present work indicate that season was the source of variation with the greatest effect on chemical characteristics of bulk-tank milk, except for casein/fat ratio for which the difference was smaller. The feeding strategy effects on characteristics of milk have also been confirmed Moreover, it has been demonstrated that the feeding strategies are applied more coherently according to their productive characteristics in farms with small and medium size than in farms with the largest herd size.

The first goal of this work was the description of a model addressed to quantify the carbon footprint in Spanish autochthonous dairy sheep farms (Manchega group), foreign dairy sheep farms (foreigners group: Lacaune and Assaf breeds), and Spanish autochthonous dairy goat farms (Florida group). The second objective was to analyze the GHG emission mitigation potential of 17 different livestock farming practices that were implemented by 36 different livestock farms, in terms of CO₂e per hectare (ha), CO₂e per livestock unit (LU), and CO₂e per liter of fat- and protein-corrected milk (FPCM). The study showed the following results: 1.655 kg CO₂e per ha, 6.397 kg CO₂e per LU, and 3.78 kg CO₂e per liter of FPCM in the Manchega group; 12.634 kg CO₂e per ha, 7.810 CO₂e kg per LU, and 2.77 kg CO₂e per liter of FPCM in the Foreigners group and 1.198 kg CO₂e per ha, 6.507 kg CO₂e per LU, and 3.06 kg CO₂e per liter of FPCM in Florida group. In summary, purchasing off-farm animal feed would increase emissions by up to 3.86%. Conversely, forage management, livestock inventory, electrical supply, and animal genetic improvement would reduce emissions by up to 6.29%, 4.3%, 3.52%, and 0.8%, respectively; finally, an average rise of 2 °C in room temperature would increase emissions by up to 0.62%.

Staphylococci are one of the main microorganisms responsible for intramammary infections in sheep, causing important economic losses for farmers and eventually health problems in humans, especially by the consumption of dairy products made with raw milk containing toxic compounds, such as biogenic amines or antibiotic resistant bacteria. This study aimed to check the presence and safety of staphylococci in bulk tank ewe’s milk from different farms, and to determine the relationship between the presence of these staphylococci and farming practices, by applying nonlinear canonical correlation models (OVERALS). Two-hundred and fifty-nine staphylococci from milk samples from eighteen farms were genotyped and representative isolates of the major clusters were identified as belonging to Staphylococcus (S.) aureus, S. epidermidis, S. arlettae, S. lentus, S. simulans, and S. chromogenes species. Identified isolates were assayed in terms of their safety, by evaluating resistance to antimicrobial drugs and the aminobiogenic capacity, using both phenotypic and genetic assays. Antibiotic resistance phenotypic assay revealed that 82.9% were resistant to some antibiotics, although in the genotypic assay only the genes tetM, ermB, ermC, and grlA were detected. Fifty-three percent were high biogenic amine (BA) producers, being putrescine the most produced amine. A lowered risk of finding antibiotic-resistant and BA-producing staphylococci is related to some farming methods such as enrolling in a breeding program, use of good farming practices, postdipping teat disinfection, hygienic livestock housing, or periodic check of the milking machine.

Adjunct cultures originating from artisanal cheese environments may play an important role in recreating and developing traditional cheese flavours, thanks to their enzymatic activities, involved in different metabolic pathways that occur during cheese ripening. In this work, Ligilactobacillus salivarius SP36, a strain isolated from an old cheese seal, was added as an adjunct culture to the cheese’s raw milk, and its effect on the microbiological, physical–chemical and sensory characteristics of the cheese was studied. The use of L. salivarius SP36 in cheese manufacturing had no significant (p > 0.05) effect on the cheese microbiota, gross composition (fat percentage, protein, total solids, moisture and NaCl concentration), colour or texture of the cheese. However, L. salivarius SP36 increased (p < 0.01) the formation of 25 volatile compounds, including 10 esters, 1 aldehyde, 8 alcohols and 6 carboxylic acids. In addition, cheeses made with L. salivarius SP36 received higher scores (p < 0.01) for aroma intensity and quality than control cheeses. L. salivarius SP36 proved to be a good candidate as an adjunct culture for cheesemaking, since it improved the cheese flavour by making it more intense and recovering typical sensorial notes of traditional cheeses.