Antimicrobial Efficacy of Cinnamon Essential Oil against Avian Pathogenic Escherichia coli from Poultry.

Colibacillosis, caused by E. coli, is responsible for economic losses in the poultry industry due to mortality, decreased production, and the cost of antibiotic treatments. Prevention of colibacillosis is based on improved biosecurity measures and the use of the vaccine performed with O78 E. coli strains, which is responsible for most cases of colibacillosis. Recently, there has been increased interest in other infection control methods, such as the use of natural compounds. The aim of this study was to evaluate the antimicrobial efficacy of cinnamon essential oil (CEO) against E. coli strains isolated from poultry. The MIC50 and MIC90 of CEO were determined by testing 117 strains belonging to serogroups O78, O2, O128, O139, isolated from laying hens (91 strains), broilers (10 strains), and turkeys (16 strains). The bacterial strains were tested at cell densities of 108 and 106 CFU/mL. At the cell density of 108 CFU/mL, MIC50 and MIC90 were 0.4 and 0.5 µL/mL for most of the tested strains, while they corresponded to 0.5 µL/mL for all strains isolated from broilers and for strains belonging to serogroup O139. At the cell density of 106 CFU/mL, MIC50 and MIC90 were 0.3 and 0.4 µL/mL, regardless of bird species of origin and for strains belonging to serogroups O78 and O2. In addition, a concentration of 0.04 µL/mL of CEO corresponded both to MIC50 and MIC90 for strains belonging to serogroups O139 and O128. Based on these results, cinnamon essential oil showed an effective antibacterial activity against E. coli strains from poultry and could find field application for the prevention of colibacillosis.

Antimicrobial Efficacy of Phyto-L, Thiosulfonate from Allium spp. Containing Supplement, against Escherichia Coli Strains from Rabbits.

Colibacillosis, caused by enteropathogenic Escherichia coli (EPEC), is one of the most common diseases in rabbit farms, resulting in economic losses due to mortality and decrease in production. Until recently, antimicrobials were used to both treat and prevent disease on livestock farms, leading to the possible risk of antimicrobial resistance (AMR) and the selection of multidrug-resistant (MDR) bacteria. Therefore, interest in alternative control methods, such as the use of natural substances, has increased in the scientific community. The aim of this study was to evaluate the antimicrobial efficacy of Phyto-L (Pro Tech s.r.l.), a product containing organosulfur compounds (OSCs) such as propyl propane thiosulfonate (PTSO) from Allium spp., against 108 strains of E. coli isolated from rabbits with colibacillosis from 19 farms. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of Phyto-L were assessed. Bacterial suspensions with a charge of 108 CFU/mL, corresponding to those found in the rabbit gut under pathologic conditions, were tested with different concentrations from 20 to 0.15 µL/mL of Phyto-L. For each strain, the MIC and concentrations above the MIC were plated on Tryptic Soy agar (TSA) without Phyto-L to assess the MBCs. MIC and MBC values ranged from 1.25 to 5 µL/mL and 1.25 to 20 µL/mL, respectively, depending on the strain tested. The data showed an interesting antibacterial activity of Phyto-L against EPEC strains. Therefore, this product could be effective in preventing colibacillosis in field application, especially considering that 104-105 CFU/g of feces is the amount of E. coli usually found in the gut contents of rabbits under physiological condition.

In Vitro Antimicrobial Effectiveness Tests Using Garlic (Allium sativum) against Salmonella enterica Subspecies enterica Serovar Enteritidis.

In recent years, there has been a more prudent use of drugs on livestock farms and alternative products have been considered, with a view of reducing the risk of the onset of antibiotic resistance. Salmonella enterica subsp. enterica serovar Enteritidis (S. Enteritidis) may cause disease in poultry, and it is also responsible for human food poisoning. The aim of this study was to evaluate the efficacy of garlic against S. Enteritidis and to define its Minimal Inhibitory Concentration (MIC)90 and MIC50 values. The study was carried out in vitro, testing 26 S. Enteritidis strains identified in laying hens from various farms in Southern Italy. A preliminary efficacy trial was carried out on two strains, S. Enteritidis and Escherichia coli, using a garlic concentration of 10 mg/mL (1%). Later, 26 strains of S. Enteritidis at 106 Colony Forming Unit (CFU) and 104 CFU were tested with different concentrations of garlic, ranging from 10 mg/mL to 1 mg/mL. Based on the results, intermediate concentrations of garlic, from 5 mg/mL to 4 mg/mL and 4 mg/mL to 3 mg/mL, were used to test 106 CFU and 104 CFU, respectively. The data were statistically analyzed. The MIC90 was 4.75 mg/mL for strains tested at 106 CFU and 4 mg/mL for strains tested at 104 CFU. The results highlight garlic's potential to inhibit the growth of Salmonella enterica ser. Enteritidis in vitro. Efficacy was dependent on the microbial concentration used. In vivo efficacy trials will be crucial to confirm the efficacy of garlic against S. Enteritidis and to assess whether garlic can be used in poultry flocks to prevent the spread of the bacterium in the field.

Prevalence and Antimicrobial Resistance of Campylobacter jejuni and Campylobacter coli from Laying Hens Housed in Different Rearing Systems.

Campylobacter (C.) jejuni and C. coli are responsible for food poisoning in humans. Laying hens may host the bacteria usually without developing symptoms. The aims of this paper were to evaluate the incidence of C. jejuni and C. coli infection in laying hen flocks housed in different rearing systems, the plasma levels of two welfare indicators (corticosterone and interleukin 6, IL-6) and the antimicrobial resistance of the detected Campylobacter strains. Two different flocks (1 and 2) from cage (A), barn (B) and aviary (C) farms were investigated. The highest (p < 0.05) levels of IL-6 were detected in laying hens housed in aviaries. A similar trend emerged in corticosterone level, although differences were found between C1 and C2. C. jejuni and C. coli were identified in 43.5% and 38.9% of birds, respectively. In total, 14 out of 177 (7.9%) hens simultaneously hosted C. jejuni and C. coli.C. jejuni was prevalently detected in hens housed in barns (B1: 53.3%; B2: 46.7%) and aviaries (C1: 34.6%; C2: 86.7%). Conversely, laying hens housed in cages were significantly exposed to infection of C. coli (A1: 41.9%; A2: 80%) while, regarding barns and aviaries, a significant prevalence emerged only in flocks B2 (40%) and C1 (54.8%). Simultaneous infection was statistically significant in barn B1 (36.7%). Antibiotic resistance was mainly detected among C. coli strains, and it was most frequent for fluoroquinolones and tetracycline. Multidrug resistance was also found in C. jejuni (19.7%) and C. coli (17.5%) strains. Based on the results of this study, we recommend increasing biosecurity and hygienic measures to manage hen flocks.

Prevalence and Antimicrobial Resistance of Campylobacter jejuni and Campylobacter coli in Wild Birds from a Wildlife Rescue Centre.

Climate change, excessive exploitation of agricultural land which reduces natural habitats, wildlife shooting, and the use of pesticides all cause difficulties for wildlife, with considerable numbers of animals being brought to wildlife rescue centres. Although the efforts of staff involved in wildlife management at these centres usually focus on therapeutic treatments to reintroduce them into the wild, the monitoring of pathogens that may be transmitted to humans is of relevance. Campylobacter (C.) jejuni and C. coli are frequently carried by animals without inducing clinical signs and are responsible for enteric disorders and more rarely extra-intestinal disease in humans. Farm species and poultry, in particular, are the main reservoirs of C. jejuni and C. coli, but wild animals may also be carriers. The aim of this paper was to investigate the presence of C. jejuni and C. coli in wild birds housed at a wildlife rescue centre and to evaluate the sensitivity of the detected strains to antibiotics. Campylobacter was found in 52 out of 209 (24.88%) birds from 33 different species. C. jejuni was more prevalent, while C. coli was only detected in three Long-eared Owls (Asio otus). The incidence of the infection was particularly high (72.22%) among omnivorous species. Infection rates were higher in birds housed indoors (57.14%) than outdoors (31.74%). Moreover, Campylobacter was not detected in species whose mean temperature body is below 40 °C or higher than 42.2 °C. The most common antibiotic resistance in the tested strains was against trimethoprim/sulfamethoxazole, ciprofloxacin and enrofloxacin. In addition, multi-drug resistance was also found. The results highlight the need to increase biosecurity measures at rescue centres so as to reduce health-related risks to workers involved in wildlife management.

New approaches for risk assessment and management of bovine protothecosis.

Protothecosis is a potential zoonosis related to bovine mastitis. In several countries, a higher incidence of protothecal bovine mastitis that is being recorded and the resistance of Prototheca species to various factors (chlorine, high temperatures, antimicrobial and antiseptic treatments, pH variations), make it difficult to control its spread among farms. The authors aim to describe the infection caused by microalgae, focusing on the problems within cattle farms and proposing new approaches to farm management, based on Regulation (EU) No 2016/429 on transmissible animal diseases. This new flexible approach, based on risk analysis, is a further tool in protecting against Prototheca species. The list of transmissible animal diseases under Regulation (EU) No 2016/429 includes those caused by microorganisms resistant to antimicrobials, which can have important implications for human and animal health, feed and food safety. This approach would involve a series of changes to the rules used for Official Controls (Regulation (EU) No 2017/625) moving from the concept of the food chain to that of the agri-food chain.

Pseudomonas fluorescens group bacteria as responsible for chromatic alteration on rabbit carcasses. Possible hygienic implications.

Bacteria belonging to the genus Pseudomonas are ubiquitous and characterized by a high adaptation capability to different environmental conditions and wide range of temperatures. They may colonize food, sometimes causing alteration. Quite recently, a blue pigmentation due to Pseudomonas fluorescens has been widely reported in mozzarella cheese. In this report, we describe a blue coloration occurred on rabbit meat stored in the refrigeration cell of a slaughterhouse. The alteration was observed after about 72 hours of storage at 4-6°C. Bacteriological analyses were performed, and a microorganism included in the Pseudomonas fluorescens group was identified. The experimental contamination was planned, using a bacterial suspension with 1×108 UFC/ml load to spread on rabbit carcasses. The blue pigmentation appeared after 24 hours of storage in a cell with the same conditions of temperature. The bacterium was reisolated and identified as responsible for the alteration on meat. These findings highlight the importance of considering the members of the genus Pseudomonas and, more specifically, of the P. fluorescens group when the microbiological quality of food is to be ascertained. In fact, even if these bacteria are not considered a public health problem, their presence should be monitored by food industry operators in self-control plans because they may cause alteration in food. In fact, any altered product should be withdrawn from the market in agreement with Regulation (EC) No 178/2002 of the European Parliament and of the Council.

Spreading of Pasteurella multocida Infection in a Pet Rabbit Breeding and Possible Implications on Healed Bunnies.

The number of pets such as dogs, cats, rabbits, and parrots has increased in European families. Social benefits to owners such as decreasing feelings of loneliness and anxiety are provided by pets which are also used in Animal-assisted Therapy (AAT). Nevertheless, human-animal interactions are also associated with health problems including allergies, asthma, and zoonosis. Rabbits may carry potential pathogens for humans. One of the most common bacteria that colonizes the oro-pharynx and the upper respiratory tract of rabbits is Pasteurella (P.) multocida. Transmission of the infection to humans results from scratches, licks, and bites but it also can occur from the inhalation of air particles containing the microorganism. Immunocompromised people or persons with pulmonary disorders are particularly susceptible to the infection. Infected rabbits may carry P. multocida with or without clinical signs. In this paper, the sensitivity to antibiotics and the invasiveness ability of P. multocida identified in a farm of pet rabbits affected by severe pasteurellosis were investigated. The strain was P. multocida belonging to capsular type A which is the type most often detected in humans. The identified strain was susceptible to the tested antibiotics, but it appeared equipped with several virulence genes which are responsible for fimbriae production, adhesion processes to host cells, enzyme production, and are involved in iron acquisition processes. These findings are of particular interest because rabbits recovered from pasteurellosis very often become carriers of the bacteria. Therefore, we suggest considering P. multocida screening in the routine medical checks of rabbits, especially if they are meant to be companion animals for children and elder people, given that the transmission of the pathogen cannot be excluded.

Religious slaughtering: Implications on pH and temperature of bovine carcasses.

The main objective of this study was to investigate the influence of different types of slaughtering on the variations of pH and temperature. Ninety entire male beef cattle, belonging to the Charolais breed, were distributed in three groups based on method of slaughtering. A total of 30 subjects (group K) were slaughtered without stunning, according to the Koscher rite; thirty bovines (group T) were stunned with a captive bolt gun, following the common slaughtering reported by Council Regulation (EC) 1099/2009; other thirty animals (group H) were slaughtered without stunning, based on Halal procedures. The temperature and pH values of carcasses were determined at 3, 6 and 24 h post-mortem, respectively. At 24 h post-mortem of the 30 carcasses evaluated for each group, muscle temperature decreased in all three types of slaughtering. In particular, the average temperature was 4.0, 4.9 and 3.0 °C for traditional, Halal and Koscher slaughtering, respectively. Moreover, at the same time step (24 h post-mortem), muscle pH decreased showing values equal to 5.72, 5.83, 5.81 for traditional, Halal and Koscher slaughtering, respectively. The findings of the present study may be useful to highlight the influence of slaughtering method on pH and, more generally, on meat quality. Additionally, our results show that, in slaughtering carried out without prior stunning, the carcass could be more susceptible to alterations because of the high pH values.

A Review on the Marek's Disease Outbreak and Its Virulence-Related meq Genovariation in Asia between 2011 and 2021.

Marek's disease is an infectious disease in poultry that usually appears in neural and visceral tumors. This disease is caused by Gallid alphaherpesvirus 2 infection in lymphocytes, and its meq gene is commonly used in virulent studies for coding the key protein functional in oncogenic transformation of the lymphocytes. Although vaccines have been introduced in many countries to control its spread and are proven to be efficient, recent records show a decline of such efficiency due to viral evolution. In this study, we reviewed the outbreak of Marek's disease in Asia for the last 10 years, together with associated meq sequences, finding a total of 36 studies recording outbreaks with 132 viral strains in 12 countries. The visceral type is the most common (13 in 16 studies) form of Marek's disease, but additional unobserved neural changes may exist. MD induces liver lymphoma most frequently (11 in 14 studies), and tumors were also found in spleen, kidney, heart, gizzard, skin, intestine, lung, and sciatic nerve. Twelve viral strains distributed in China have been reported to escape the CVI988 vaccine, reaching a mortality rate of more than 30%. Phylogenetic analyses show the internal connection between the Middle East (Turkey, Iraq, Iran, Saudi Arabia), South Asia (India, Indonesia), and East Asia (China and Japan), while external viral communications might occasionally occur. In 18 strains with both sequential and mortality data, amino acid alignment showed several point substitutions that may be related to its virulence. We suggest more behavioral monitoring in Marek's disease-endemic regions and further studies on strain virulence, together with its Meq protein structural changes.

Salmonella Infection in Turtles: A Risk for Staff Involved in Wildlife Management?

Monitoring of infections that may be transmitted to humans by animals in wildlife rescue centres is very important in order to protect the staff engaged in rehabilitation practices. Salmonella may be a natural inhabitant of the intestinal tract of turtles, rarely causing disease. This may represent a potential risk for humans, increasing the sanitary risk for operators in wildlife rescue centres. In this paper, the occurrence of non-typhoidal Salmonella among terrestrial turtles housed in a wildlife rescue centre in Southern Italy was investigated, in order to assess the serovars more frequently carried by turtles and identify those that may represent a risk for operators involved in wildlife management. Sixty-nine adult turtles (Testudo hermanni hermanni, T. h. boettgeri, T. graeca, and T. marginata) were tested. Detection and serotyping of Salmonella strains were performed according to ISO 6579-1 and ISO/TR 6579-3:2013, respectively. The distribution of Salmonella spp. was significantly higher in T. hermanni hermanni than in other species, independent of the age and gender of the animals. Two different Salmonella species, S. enterica and S. bongori, three S. enterica subspecies (enterica, diarizonae, salamae), and five different serovars (Hermannswerder, Abony, Ferruch, Richmond, Vancouver) within the group S. enterica subspecies enterica were identified. Different combinations of Salmonella types were simultaneously found in specimens of T. h. hermanni. Most of detected Salmonella types may represent a potential risk for public health. Adopting correct animal husbandry procedures and informing on potential sanitary risks may be useful for minimising the risk of transmission of Salmonella to workers involved in wildlife management.