Using Expert Elicitation for ranking hazards, promoters and animal-based measures for on-farm welfare assessment of indoor reared beef cattle: an Italian experience.

On-farm welfare assessment gives the opportunity to monitor and improve the quality of the animal life on the farm. In order to build the first Italian public standard for the welfare of indoor reared beef cattle, a list of 25 potential hazards and 22 potential promoters of beef cattle welfare was judged by a group of national experts by taking into account their negative or positive impacts on the welfare-state of the target population. In addition, the experts were asked to rank a list of 11 animal-based measures for identifying the most appropriate and important for measuring negative welfare outcomes in beef cattle. Based on experts' ratings, an "impact score" (ISoverall) was calculated for the proposed measures. Management hazards and promoters were ranked to have a greater impact on beef cattle welfare than housing factors. Keeping cattle in large (≥ 40 animals) and heterogeneous groups obtained the highest ISoverall among the proposed hazards (ISoverall = 5.54), followed by the presence of animals without free access to drinking water (ISoverall = 4.39) and the use of high-concentrate corn silage diets (concentrate > 80% and fiber < 6%) (ISoverall = 4.39). On the other hand, housing animals in small (≤ 20 animals) and homogeneous groups (ISoverall = 5.41), checking them at least twice a day (ISoverall = 4.62) and rearing cattle in loose housing systems with access to an outdoor area/pasture (ISoverall = 4.27) were ranked among the top 3 promoters. Concerning animal-based measures, experts scored lameness, severe respiratory diseases, body condition scoring and mortality rate to be measures most important for assessing serious welfare impairment.

Promoting Judicious Antimicrobial Use in Beef Production: The Role of Quarantine.

Judicious antimicrobial stewardship in livestock industry is needed to reduce the use of antimicrobials (AMU) and the associated risk of antimicrobial resistance. Biosecurity measures are acknowledged for their role against the spread of diseases and the importance in reducing AMU in different species. However, their effectiveness in beef production has been scarcely considered. The aim of this study was to investigate the effect of the quarantine strategy on AMU in beef cattle. A total of 1206 Charolaise animals in five farms were included in the trial. Roughly half of the animals followed the standard procedure of the fattening cycle (no-quarantine; NO-QUA group) and half followed a 30-day period of quarantine (QUA group) since their arrival. Performance and antimicrobial data were recorded and a treatment incidence 100 (TI100it) per animal was calculated. Penicillins was the most used class of antimicrobials. Differences between groups were significant for males only, with NO-QUA group having greater TI100it (3.76 vs. 3.24; p < 0.05) and lower body weight at slaughter (713.4 vs. 723.7 kg; p < 0.05) than QUA group. Results suggest that quarantine strategy can reduce AMU in males without compromising their performance, whereas further investigation is needed for females.

The influence of different production systems on dioxin and PCB levels in chicken eggs from Emilia-Romagna and Lombardy regions (Italy) over 2017-2019 and consequent dietary exposure assessment.

Hen eggs from farms with different production systems (organic, free range, barn, and in cage) sited in two regions of Italy (Emilia-Romagna and Lombardy) were collected from 2017 to 2019 to monitor the levels of 17 polychlorinated dibenzo-p-dioxin/dibenzofurans (PCDD/Fs), 12 dioxin-like polychlorinated biphenyls (dl-PCBs) and the 6 non-dioxin-like polychlorinated biphenyl indicators (ndl-PCBs). Average concentrations of PCDD/Fs (0.21 pg WHO(2005)-TEQ/g fat), PCDD/Fs + dl-PCBs (0.43 pg WHO(2005)-TEQ/g fat) and ndl-PCBs (6.41 ng/g fat) were below the maximum limits established at European level in eggs, but significantly higher amounts of PCDD/Fs and PCDD/Fs + dl-PCBs were found in eggs from free-range housing system compared to barn, cage, and organic ones. The potential dietary intake of the monitored contaminants by Italian population age groups through the consumption of locally produced eggs was also evaluated. Exposure levels to PCDD/Fs plus dl-PCBs were below the tolerable weekly intake (TWI) of 2 pg WHO(2005)-TEQ/kg bw/week recently set by the European Food Safety Authority (EFSA). However, the younger population was found to be more vulnerable than other age groups to exposure to these contaminants. Specifically, the preferential consumption of free-range eggs by infants, toddlers, and children contributed more than 30% to the TWI. The results confirm the need to continuously monitor the levels of chemical contaminants in the environment and provide a reminder of the importance of targeted intervention measures aimed to reduce their occurrence in food and feed, firstly by improving the management practices on egg-producing farms.

The Relationship between Animal Welfare and Antimicrobial Use in Italian Dairy Farms.

Information regarding the relationship between animal welfare (AW) and antimicrobial use (AMU) in dairy cows is limited. The current study aimed to investigate this relationship on Italian farms and to identify potential targets of AMU reduction. The study was performed at 79 Italian dairy farms housing over 15,000 cows during 2019. AW was scored with an on-farm protocol assessing farm management and staff training, housing systems, and animal-based measures. AMU was estimated using a defined daily dose per kg of animal biomass (DDDAit/biomass) for Italy. The median AW score was 73% (range: 56.6-86.8%). The median AMU was 4.8 DDDAit/biomass (range: 0-11.8). No relationship between the total AMU and AW was found. Management and staff training were positively associated with the use of the European Medicines Agency's category B antimicrobials, which are critical for human medicine, and with intramammary products for dry cow therapy. In those farms, antimicrobial stewardship should aim to reduce the category B antimicrobials and selective dry cow therapy. Our results underline the importance of implementing both an integrated monitoring system (AW, AMU, etc.) and antimicrobial stewardship tailored to the specific needs of each dairy farm.

Best Farming Practices for the Welfare of Dairy Cows, Heifers and Calves.

The concept of animal welfare (AW) has many meanings. Traditionally, AW has been considered as freedom from disease and suffering. Nowadays, growing attention goes to the concept of "positive animal welfare" (PAW), which can be interpreted within the concept of quality of life (QoL), thinking about a "balance of positives over negatives" and a "life worth living". In this vision, where the QoL represents a continuum between positives and negatives, the Italian National Reference Centre for Animal Welfare (CReNBA), within the Istituto Zooprofilattico Sperimentale della Lombardia e dell'Emilia Romagna (IZSLER), has developed a welfare assessment protocol for dairy cows, heifers, and calves in loose housing systems, including both animal-based and non-animal-based indicators, in which not only hazards but also benefits are identified. This protocol is part of an integrated monitoring system called "ClassyFarm", belonging to the Italian Ministry of Health and developed by IZSLER. The aim of this paper is to extrapolate from the mentioned protocol, a list of 38 best farming practices (on managerial and equipment factors) for ensuring a high level of welfare in dairy cattle. All stakeholders (veterinarians, farmers, competent authorities, consumers, etc.) can benefit of these best practices as a guide or toolbox to ensure a life worth living for these animals.

Welfare Assessment: Correspondence Analysis of Welfare Score and Hematological and Biochemical Profiles of Dairy Cows in Sardinia, Italy.

The need for animal welfare definition and assessment is increasing worldwide, and several studies have been conducted to help fill the knowledge gaps regarding the welfare of cattle. However, further studies are needed to provide valid synthetized measures for welfare evaluation. The aim of this study was to assess the welfare status of 16 Sardinian dairy cattle farms, based on the developed Animal Welfare and Biosecurity Evaluation checklist (AWB-EF) and the corresponding hematological, biochemical, and electrophoretic profiles of these animals. Considering the AWB-EF as gold standard, blood samples were collected from 230 Holstein breed dairy cattle, aged between 3 and 8 years, out of the periparturient period, and with no clinical signs of specific pathologies. Principal Component (PC) and correlation analyses were performed to simplify phenomena interpretation and assess positive/negative associations. Four PCs were able to explain 76% of the total variability, and six laboratory parameters were strongly associated with the AWB-EF score (Spearman's correlation coefficient ≥ 0.40, p-Value < 0.05), reflecting the real health status of the animals. Given the complexity of animal welfare as a multidimensional concept and the need to include both animal-based and non-based measures in welfare evaluation, the present work represents a sound basis for future evaluation and veterinary health planning.

Effect of welfare standards and biosecurity practices on antimicrobial use in beef cattle.

Antimicrobial use (AMU) in livestock species and the associated antimicrobial resistance are a global concern, thus strategies for their reduction and a more judicious use are needed. Previous research has revealed a link between improved animal welfare, biosecurity and AMU reduction in pig and dairy sectors, however, little is known about the beef sector. This study aimed to investigate the impact of welfare standards and biosecurity on AMU in beef cattle. Data on performance traits and AMU were collected over a 3.5 year time from 27 specialised beef farms and a treatment incidence was calculated using the defined daily dose for animals. An on-farm assessment was carried out by assigning a score from 0 (very poor) to 100% (very good) to 3 sections: welfare, biosecurity and emergency management. The highest average score was obtained for the welfare section (76%) followed by emergency management (39%) and biosecurity (24%). This suggests that major focus on strategies for the implementation of biosecurity measures and emergency management is needed, due to the low scores reported. A statistically significant lower AMU was observed with improved level of welfare. These results may be helpful for farm benchmarking and highlight the importance of improved animal welfare for an efficient antimicrobial stewardship.

Use of antimicrobials in beef cattle: an observational study in the north of Italy.

Antimicrobial resistance is a worldwide issue whereby a more prudent use of medications is needed, especially for those antimicrobials (AM) classified as 'highest priority critically important antimicrobials' (HPCIAs) which are likely contributors to the development of resistance. So far, data on antimicrobial use (AMU) in EU are mainly reported at sales level while information on real use, mostly in beef production, is poor. The most reliable indicator to measure AMU is the treatment incidence (TI100) calculated by using the Defined Daily Dose Animal (DDDA) as stated by the European Medicines Agency (EMA). Although Italy ranks second among EU countries with regard to the AM sales in livestock production, data on AMU of the Italian beef production is still lacking, whereby the aim of this study was to provide information on the current scenario of AMU in Italian beef cattle. Data were collected from January 2016 to April 2019 from specialized beef fattening farms located in the north of Italy yielding a final dataset of 1376 batches. Data on performance and AM agents used in the study were collected and TI100 indexes per batch were calculated according to both Italian and EMA's DDDA. A Kruskal-Wallis test was used to check for differences between years and seasons. Results showed a significant variation between years and seasons with a reduction of TI100 based on Italian DDDA as time progressed (P < 0.05). However, about 40% of the total amount of treatments administered were HPCIAs with macrolides accounting for the 27.7% of the total amount. The most common reasons of administration of AM were respiratory diseases (68.9%) and lameness (17.6%). Penicillins was the class of AM used on the highest proportion of batches (84.4%) showing that broad-spectrum AM were widely exploited among herds. In summary, despite a general reduction of AMU in beef cattle over time, a great use of HPCIAs was still observed suggesting that AM stewardship for Italian beef production should pay particular emphasis on the reduction of HPCIAs use. This shows how overall knowledge on where efforts need to be optimized is important to develop targeted strategies for a more responsible AM stewardship. Results of the current study may also contribute to define national and EU benchmark criteria for AMU, as a comparison with studies carried out in other countries or on other food-producing sectors is still challenging to achieve.

Bulk tank milk quality data is unlikely to give useful information about dairy cow welfare at herd level.

This research communication explores the value of routinely collected bulk tank milk quality data for estimating dairy cattle welfare at herd level. Selected bulk tank milk quality parameters (somatic cell count, total bacterial count, urea, protein and fat contents) recorded during the years 2014-2016 in 287 Italian dairy farms were compared with the animal welfare data of each farm. The welfare assessment data were extracted from the database of the Italian Reference Centre for Animal Welfare (CReNBA), which includes the outputs of the application of the CReNBA welfare assessment protocol for dairy cows, used at national level for on-farm controls. The statistical analysis was carried out using the correlation coefficient for Kendall's Tau ranks, in order to investigate the presence of a categoric relationship between the selected bulk tank milk quality parameters and the overall animal welfare score or the scores of the single areas A (farm management and staff training), B (housing) and C (animal-based measures). Somatic cell count, total bacterial count, urea and proteins demonstrated only a few statistically significant and very weak correlations with farm animal welfare data, while no significant correlations were obtained for milk fat content. Given the weak correlations found, the selected bulk tank milk parameters seems to be able to provide only limited information about the welfare level of the herd, thus it could be difficult to use them for drawing up a pre-screening model for identifying herds at risk of poor welfare.

PCDD/Fs, DL-PCBs, and NDL-PCBs in Dairy Cows: Carryover in Milk from a Controlled Feeding Study.

A feeding study was carried out to investigate the kinetics in cow milk of the 17 polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDD/Fs), the 12 dioxin-like polychlorinated biphenyls (DL-PCBs), and the 6 non-dioxin-like PCBs (NDL-PCBs) regulated by the European (EU) legislation. A fortified ration (ΣPCDD/Fs and DL-PCBs: 24.68 ng TEQ/day/cow; ΣNDL-PCBs: 163.99 µg/day/cow) was given to the animals for 49 days, followed by 42 days on clean feed. EU maximum limit for TEQPCDD/F+DL-PCB was exceeded in milk after 1 week of exposure, while for ΣNDL-PCBs, after 5 weeks. Milk compliance was restored after 1 week on clean feed, but to return to the basal TEQPCDD/F+DL-PCB it took 42 days. At the end of the study, ΣNDL-PCBs had not yet reached the basal level. The carryover rate of ΣNDL-PCBs was 25.4%, while the carryover rate of TEQPCDD/F+DL-PCB was 36.9%. The latter was mainly affected by the 12 congeners contributing most to the toxic equivalent (TEQ) level, explaining the fast overcome of the maximum limit in milk.

Diagnostic value of composite milk sample vs single quarter milk sample for the detection of Staphylococcus aureus intra-mammary infections in cattle.

Staphylococcus aureus (S. aureus) is one of the most important causes of mastitis in dairy cattle. Control and eradication programs of S. aureus intra-mammary infections (IMI) are based on different factors included the correct detection and management of the infected cows. The present study aimed at evaluating the efficacy of composite milk samples (CMS) analysis, compared to quarter milk samples (QMS) analysis, for the bacteriological detection of S. aureus intra-mammary infections. During 2016, 661 CMS (hygienically collected) and 2644 QMS (aseptically collected) were obtained from 661 cows in 5 herds. All the samples were submitted to S. aureus bacteriological culture and somatic cell count (SCC) analysis. QMS bacteriological analysis on blood agar plates was able to detect 236 cows excreting S. aureus, while the bacteriological analysis of CMS, using selective agar, identified 229 positive cows. The concordance was 95% with an excellent Cohen's κ (0.89). Relative sensitivity and specificity of CMS vs QMS, considered as the reference test, were 91.5% ± 2.1 and 96.9% ± 1.3 (CI 95%), respectively. In addition, the relative sensitivity of CMS improved as the number of infected quarters per cow and the number of colony forming units (cfu) per sample increased. The predictive value of CMS results was better when paired with SCC data, in particular CMS showed better negative predictive value when SCC was <200,000 cells/mL and better positive predictive value when SCC was>200,000 cells/mL. The probability for a cow to be S. aureus positive was 56.4% in case of SCC > 200,000 cells/mL, while it was 18.6% in case of SCC < 200,000 cells/mL. The average SCC in CMS was significantly higher in positive cows and the value rose as the number of infected quarters per cow increased. Given the intermittent excretion of S. aureus in milk from dairy cows, it could be more advantageous to carry out several serial CMS, rather than few QMS, being CMS an easier to collect and less expensive milk sampling method. Thus, bacteriological examination of CMS, combined with SCC data of the same sample, could be extremely useful for the success of S. aureus IMI control plans, because repeated CMS are easier to be performed and could be more easily proposed to the farmers.

Characterization of hazards, welfare promoters and animal-based measures for the welfare assessment of dairy cows: Elicitation of expert opinion.

An expert opinion elicitation, based on a modified Delphi technique, was organized to collect the opinion of 16 Italian veterinarians with the aim of conducting a hazard and a welfare promoter characterization for defining and weighing a list of management and housing factors potentially associated with negative or positive welfare outcomes in dairy cows kept in loose housing systems. In addition, the 16 experts judged a set of animal-based measures in order to rate them by appropriateness and by the level of animal pain and suffering due to the welfare consequences they measure. Veterinary experts were asked to score 52 hazards, 47 welfare promoters and 18 animal-based measures. Management and housing hazards, that were determined to be associated with a very high impact on the welfare of dairy cows, were mainly referred to lactating cows (absence of bedding material, presence of inadequate or slippery floor in walking areas, wrong design of the lying area), but also the use of harmful tools for animal handling and the lack of scheduled foot inspection, trimming and foot bathing reached very high impact scores. Management and housing welfare promoters dealing with optimal cow comfort around resting obtained the highest ratings, together with the presence of experienced and trained stockpersons, the implementation of an attentive animal grouping strategy and the control of temperature, humidity and ventilation. Considering animal-based measures, the highest ranking of importance was given to observation of lameness, records of mortality of adult cows and calves, observation of integument alterations and body condition scoring. This study has been the starting point for the development of the first Italian national protocol for the assessment of the welfare of dairy cows farmed in loose housing systems.

Assessing circumstances and causes of dairy cow death in Italian dairy farms through a veterinary practice survey (2013-2014).

A questionnaire survey about on farm dairy cow mortality was carried out among veterinary practitioners in Italy between January 2013 and May 2014. The study aimed at investigating the main circumstances of death in dairy cows (euthanasia, emergency slaughter or unassisted death), the primary causes and the risk factors of death. Out of 251 dead cows involved (across 137 farms), 54.6% died assisted and 45.4% were found dead. The main causes of death were metabolic/digestive disorders (22.3%) and mastitis/udder problems (17.1%), while in 14.7% of all cases, reasons of death were unknown. From the univariable generalised linear mixed models, dry cows showed a significantly higher odds to die unassisted compared to lactating cows (OR=3.2); dry cows also had higher odds of dying from unknown reasons (OR=11.7). Season was not significantly related to the risk of dying unassisted and for unknown reasons, but during the summer (characterised by hot and muggy weather in Northern Italy) cows died mostly for problems at calving. 54.2% of cows died during the first 30days in milk (DIM). Half of the multiparous cows that died, died in the first 29.5 DIM, while half of the primiparous cows that died, died in the first 50 DIM. Results pointed out that, especially in dry cows, around calving and during the summer, some failure in management practices and daily inspections may occur. Improvements should be done in monitoring activities and in recognising early symptoms of diseases among stockperson. In addition, in case of diagnosed diseases with poor prognosis, euthanasia procedures should be implemented to prevent cows from dying unassisted.

Case-study and risk management of dioxins and PCBs bovine milk contaminations in a high industrialized area in Northern Italy.

Milk supplied to a dairy plant in Brescia City (Northern Italy) was found to be contaminated by dioxin like PCBs at levels above the European (EU) action limit (2 pg WHO-TEQ/g fat). As a consequence, 14 dairy farms were sampled individually, in order to identify and possibly eliminate the source of contamination. All the farms were located in Brescia or just nearby, an area that is characterized by a strong industrialization. Four out of the 14 farms showed contamination levels above the legal maximum limit set by European Commission at 5.5 pg WHO-TEQ/g fat for the sum of dioxins and DL-PCBs. Concentrations of 8.16, 6.83, 5.71 and 5.65 pg WHO-TEQ/g fat were detected. In the three most polluted farms, cow ration was substituted with feed coming from uncontaminated areas and the time needed to reduce milk pollution was evaluated. In all the three farms, contamination levels dropped below the EU legal limit after only 1 month from the removal of the pollution source. In each sampled farm, DL-PCBs were the major contributors to the total WHO-TEQ level, with percentages up to 87% in the most contaminated one. PCB 126 WHO-TEQ value explained by itself large part of this contamination, and its decrease was fundamental for the reduction of milk contamination levels. This study provides an example of an on-field successful emergency intervention that succeeded in decontamination of dairy cows, allowing a fast restart of their production activity.

Aflatoxin control in maize by Trametes versicolor.

Aspergillus flavus is a well-known ubiquitous fungus able to contaminate both in pre- and postharvest period different feed and food commodities. During their growth, these fungi can synthesise aflatoxins, secondary metabolites highly hazardous for animal and human health. The requirement of products with low impact on the environment and on human health, able to control aflatoxin production, has increased. In this work the effect of the basidiomycete Trametes versicolor on the aflatoxin production by A. flavus both in vitro and in maize, was investigated. The goal was to propose an environmental loyal tool for a significant control of aflatoxin production, in order to obtain feedstuffs and feed with a high standard of quality and safety to enhance the wellbeing of dairy cows. The presence of T. versicolor, grown on sugar beet pulp, inhibited the production of aflatoxin B1 in maize by A. flavus. Furthermore, treatment of contaminated maize with culture filtrates of T. versicolor containing ligninolytic enzymes, showed a significant reduction of the content of aflatoxin B1.

Investigation on the origin of prednisolone in urine and adrenal glands of cows.

Prednisolone is a steroid belonging to the corticosteroid group. The results obtained in the application of the 2008 and 2009 Italian Residue Control Plans show the frequent detection of prednisolone traces in cow's urine. Since most of the positive samples were detected at the slaughterhouse, the researchers hypothesised that, together with an increase of cortisol concentration, traces of prednisolone could be produced endogenously during stressful situations due to transport and handling before slaughter. In the present trial, 52 lactating cows housed in seven different farms in Lombardy, Italy, were studied. Urine samples were collected at the farm (after urethral catheterisation) and immediately after slaughter (from urinary bladder) together with 40 adrenal gland samples belonging to the same animals. All the samples were analysed for the determination of prednisolone and cortisol by LC/MS(n). The results demonstrated that prednisolone can be endogenously produced in dairy cows and, furthermore, its endogenous presence in bovine urine seems to be strongly related to a state of stress in the animals (at the farm and at the slaughterhouse). The data from adrenal glands do not, however, clarify if the endogenous production occurs, partially or totally, in this organ.