Dpto. Producción y Sanidad Animal, Salud Pública Veterinaria y Ciencia y Tecnología de los Alimentos

Introduction: Learning molecular genetics techniques is part of the program of the genetics course of the veterinary degree. It is essential to maintain motivation and interest in genetics practice by using a common thread that connects with the students’ interests. This work describes the design, implementation, and results of a gamification strategy developed during two practical sessions in which the student becomes a geneticist to identify which dog a faecal sample collected from the street belongs to, as some municipalities currently do to promote hygiene and public health in their streets. The aim was for students to understand basic concepts and techniques in molecular genetics during these two sessions. This included learning how to extract DNA from different types of samples, describing the amplification using Random Amplification Polymorphic DNA (RAPD), and getting a better understanding of molecular markers and the theory behind Polymerase Chain Reaction (PCR). Methodology: In the first practice, students extracted DNA from fresh dog faeces, while also discussing various DNA sources and extraction methods. Then they used the extracted DNA to create a simulated database of fictional dogs associated with students. They quantified the DNA, analysed its quality, and prepared a dilution to 10ng/μL. In the second practice, the students used RAPD to identify individuals by matching DNA from a simulated collected faeces sample to their fictional dogs DNA database. They performed amplification reactions with various primers pairs, followed by gel electrophoresis, to compare DNA band patterns and identify the dog and the fictional student associated with the uncollected dog faeces. The advantages and limitations of the RAPD technique were discussed, along with its potential applications in veterinary science and genetics. Results and discussion: The students were successful in extracting DNA with concentrations over 100 ng/μL in most cases as well as a good purity with respect to proteins. However, it was found that there was usually a low quality of DNA with respect to salts, although this did not influence the results of the second practice. They were able to generate reproducible RAPD profiles with all primer pairs. The unknown individual could be easily recognized within the database. Conclusions: It is concluded that this educational proposal is an effective option for teaching DNA extraction and the RAPD technique, as well as for many molecular genetics terms and concepts and contributes to the comprehensive training of future veterinary professionals. Additionally, the educational and social value of the practices are highlighted, as they promote interest in science, respect for the environment, and civic responsibility.

European hedgehogs (Erinaceus europaeus) are nocturnal insectivores frequently found in urban areas. In the last decades, their population has declined in various European countries and human activities have emerged as significant contributors to this trend. While the literature has mainly focused on trauma as the major cause of mortality, few authors have considered pathological findings. The present study is based on the results of full post-mortem examinations performed on 162 European hedgehogs in Italy and 109 in Switzerland. Unlike in previous studies, the main cause of mortality was infectious diseases (60.5%), followed by traumatic insults (27.7%). The lungs were the main organ affected, showing mostly lymphoplasmacytic (45.9%), granulomatous (18.1%) or suppurative (8.2%) pneumonia. Nematodes were detected in 57.2% of all lungs and were significantly associated with pneumonia (p-value < 0.001). To our knowledge, this is the first study to report infectious diseases as the main cause of hedgehog death, emphasizing the need for wildlife rescue centers to adopt appropriate diagnostic and therapeutic measures. Further research is necessary to determine the broad range of infectious agents that affect this species and elucidate their interplay with the host. Finally, citizen sensitization should be implemented to promote responsible behaviors that could reduce human-related traumatic events.

Since its introduction in Spain in 2004, Aedes albopictus has rapidly spread across the country. Its aggressive biting behaviour causes nuisance, limiting outdoor activities. Also, its role as a vector of several arboviruses implies a major public health risk, with several cases of autochthonous dengue having been reported nationwide over the past few years. Control strategies usually focus on interventions in breeding sites. As such, accurate knowledge of the main larval habitats becomes a major priority in infested areas. A detailed identification of breeding sites of Ae. albopictus was carried out in the outdoors of 60 residential properties during July–August 2022 in El Vedat de Torrent (Valencia, Eastern Spain), an area recently colonised by this species. A total of 1444 real and potential breeding sites were examined. The most abundant potential larval habitat were plant pot plates (6.48 units/house), although a low infestation level was found, both for larvae (2.06% positivity, x̄ = 30.5 larvae/container), and pupae (0.51%, x̄ = 2.5 pupae/container). A total of 7715 larvae and 205 pupae were found in a disused flooded water pool depuration system. Animal drinkers, buckets and irrigation water containers were found to be the most common positive containers. No statistical difference was observed among the different container materials. A general statistical increase of 1 larva per 11.7 ml of water in breeding sites was detected. Breeding sites of other species such as Culex pipiens (n = 2) and Culex modestus (n = 1) were also rarely found in this residential area. To our knowledge, this is the first aedic index study carried out in Europe, and it provides valuable information about the main domestic breeding habitats of Ae. albopictus, which can greatly improve control programmes.

Aedes albopictus was first detected in the province of Valencia, Eastern Spain, in 2013. Since then, this invasive mosquito species has become a major pest in many municipalities due to its annoyance, but also to its importance as a public health concern as a potential vector of several arboviruses. Within the framework of the NESCOTIGER research project, several prevention and control activities were carried out during spring 2022 in the residential area of Torrent, a municipality recently invaded by the Asian tiger mosquito. An awareness campaign was conducted in the study area focused to both primary school students and residents. For children, talks in primary schools focusing on the basic aspects of the biology, prevention, control, and disease transmission of Ae. albopictus were conducted. For adults, a multi-pronged education campaign to mobilize the community in response to this mosquito species was devised, involving a series of workshops and local gatherings to raise awareness about Ae. albopictus. Additionally, a knowledge, attitude and practices (KAP) study was conducted on both children (n = 544; 75.7%) and adults (n = 175; 24.3%). Both adults and children showed low levels of knowledge concerning Ae. albopictus biology, control strategies, and breeding sites identification. Nevertheless, there was great interest in learning and participating in novel strategies against this Aedes invasive mosquito. Future awareness campaigns in the area should consider the information gathered here for the development of community-based strategies. This study is the first to collect information on the KAP of local community members since the detection of Ae. albopictus in Spain, and it is the first to compare KAP surveys between adults and children concerning this invasive mosquito.

The objective of this study was to evaluate colostrum IgG concentration harvested at first and second milking from multiparous Jersey cows, the dam's lactation number, colostrum yield, and time of first milking. In addition, we validated the use of a Brix refractometer to estimate IgG concentration in colostrum from multiparous Jersey cows using radial immunodiffusion as the reference method. Colostrum samples and total weight of colostrum harvested at first (n = 134) and second (n = 68) milking were collected from 134 multiparous Jersey cows housed in a California herd. Fresh colostrum samples were analyzed for IgG concentration with Brix refractometry and frozen samples by radial immunodiffusion. A total of 90.4 and 42.7% of the samples from first and second milking met industry standards of quality for IgG concentration (>50 g/L). Second and third lactation cows had similar colostrum IgG concentration but lower than cows on their fourth and greater lactation. At second milking, 56.4% of cows on their fourth or greater lactation had colostrum IgG concentrations >50 g/L. When colostrum yield increased from low (<3 kg), medium (3 to 6 kg), to high (>6 kg), IgG concentration decreased. Higher IgG concentration was observed on colostrum harvested at <6 h (short) versus 6 to 11 h (medium) after calving. However, IgG concentration in colostrum harvested after 11 h (long) was similar to that harvested at short and medium time. Readings of %Brix were highly correlated with IgG at first (r = 0.81) and second (r = 0.77) milking. The best Brix threshold to identify colostrum from first milking with >50 IgG g/L was 20.9% based on logit equations with Youden's index criterion and 18.0% based on accuracy criterion. For colostrum harvested at second milking, similar Brix thresholds were obtained, 19.2 and 19.0%, regardless of whether Youden's index or accuracy was used as the selection criterion. Our results indicate that the dam's lactation number, colostrum yield, and time of first milking relative to calving are associated with IgG concentration in colostrum from multiparous Jersey cows. Second milking colostrum from mature Jersey cows should be evaluated to extend colostrum supply on dairies especially during times of shortage. Readings of %Brix can be used to rapidly estimate IgG concentration in Jersey colostrum harvested at first and second milking.

Indirect assessment of metabolic status using milk samples provides a non-invasive and objective tool for cow-level health monitoring. Milk fat-to-protein ratio (FPR) has been commonly evaluated as an indirect measure for negative energy balance (EB) in confined dairy cows. However, milk component ratios have not been explored for their association with pasture-based cows' metabolic status. The objectives of this observational study were to 1) describe milk component ratios from 0 to 45 d postpartum, 2) evaluate the associations between milk component ratios [FPR, fat-to-lactose (FLR), protein-to-lactose (PLR)] and indicators of EB (serum β-hydroxybutyrate (BHB) concentration at 5–45 d postpartum and body condition score (BCS) change during the transition period), and 3) evaluate the associations between milk component ratios and serum Ca concentration 0–4 d postpartum in spring-calving dairy cows from pasture-based commercial farms. Milk component ratios were determined on samples collected before AM or PM milkings from 548 cows at 0–45 d postpartum (n = 970). Serum BHB and Ca determinations were performed in blood samples collected at the time of milk sample collection at 5–45 d postpartum (n = 918) and 0–4 d postpartum (n = 50), respectively; and BCS change was calculated using BCS assigned between 29 d prepartum and 45 d postpartum (n = 851). Cows' calving date, parity (1st, 2nd–3rd or ≥ 4th) and breed (Holstein-Friesian or dairy crossbred) information was obtained from the farm records. Data was analyzed by multiple linear regression. Average milk FPR, FLR and PLR were 0.70, 0.53 and 0.72, respectively. Milk FPR linearly increased while milk FLR linearly decreased postpartum both at a rate of 0.004 units per day; milk PLR decreased 0.05 units per day for the first 30 d postpartum and moderately increased afterward. Milk FPR and FLR were 0.71 and 0.52 units lower before AM than PM milking, respectively; while milk PLR was similar before AM and PM milking. Milk FPR and FLR were 0.07 to 0.10 units higher for 2nd–3rd compared with 1st and ≥ 4th parity cows. Milk PLR was 0.03 units greater for ≥ 4th compared with 2nd–3rd and 1st parity cows. Further, crossbred cows had 0.07, 0.08 and 0.03 higher milk FPR, FLR and PLR than Holstein-Friesian cows, respectively. Moderate to high P-values along with moderate to small estimated slopes and wide 95% confidence intervals were observed for the associations between milk component ratios and indicators of EB. A positive linear association was observed between milk FPR and serum Ca concentration within 4 d postpartum; milk FPR increased 0.31 units per each mmol/L increase in serum Ca concentration. Cows with low serum Ca concentration within 4 d postpartum had 0.27 units lower milk FPR compared with cows at or above the threshold (2.12 mmol/L), and tended to have 0.15 units lower milk FPR compared with cows at or above the threshold (2.00 mmol/L). In conclusion, further research is needed to reach conclusions on the association between milk component ratios determined before milking and EB indicators. The potential of milk FPR for monitoring blood Ca status warrants further investigation in early-lactation pasture-based dairy cows.

The use of human milk in milk banks requires thermal processing to eliminate microbiological hazards. An evaluation is made of the stability of overall human milk bactericidal capacity following 2 modalities of thermal pasteurization: 63°C/30 minutes and 75°C/15 seconds. Ten milk samples (mature milk) were analyzed. In each sample, the effect of both thermal treatments on bactericidal capacity against Escherichia coli was evaluated in relation to the capacity of fresh milk (control). All the samples analyzed possessed bactericidal capacity. Human milk pasteurization induced a significant loss of this capacity that was more pronounced after high-temperature treatment than after low-temperature processing. Untreated milk, low-pasteurized milk, and high-pasteurized milk yielded a reduction in E. coli growth of 70.10%, 52.27%, and 36.39%, respectively. In conclusion, human milk possesses antimicrobial activity that is lost in part as a result of thermal processing. Such bactericidal capacity is, moreover, better preserved by low-temperature, long-time pasteurization.

The population dynamic of constitutive biota on 84 samples belonging to two different types of French fermented dry sausages during the ripening process in a pilot-scale ripening chamber was investigated. Samples were analyzed in three steps of their production: fresh product, first drying stage, and finished product. In addition, 180 strains of lactic acid bacteria were identified using a miniaturized biochemical procedure of characterization. In general, the number of lactic acid bacteria that evolved during the ripening process of French dry sausages increased during the first days of the process after which the number of these organisms remained constant at approximately 8 log CFU/g. Lactobacillus sakei and Pediococcus pentosaceus, bacteria added as starter, were the dominant species. Pediococcus urinaeequi, Pediococcus acidilactici, and particularly Lactobacillus curvatus were also present. Finally, we have to take into account that the controlled conditions of the pilot plant generally contribute to the homogenization of the behavior of the starter biota.

The individual and combined antilisterial efficiency of nisin, lactic acid and modified atmosphere packaging (MAP) was investigated. Raw ground pork was inoculated with a strain of Listeria monocytogenes and samples were distributed into twelve lots. Half of the lots were stored aerobically and the other six lots were packaged using the MAP. A different combination of nisin (N) and/or lactic acid (LA) was added to each lot (300 ppm N, 500 ppm N; 2% LA; 300 ppm N and 2% LA; 500 ppm N and 2% LA). All samples were stored at 4 °C for 21 days (samples with MAP) or 7 days (samples stored aerobically). The inactivation of L. monocytogenes in samples stored aerobically was attained mainly with the combination of 500 ppm N + LA; however, in samples with MAP, the L. monocytogenes population decreased 3.45 log with the addition of LA, and the combination N + LA increased the inactivation other 0.5 log.