Rapid detection and quantification of plasmid-mediated colistin resistance genes (mcr-1 to mcr-5) by real-time PCR in bacterial and environmental samples.

AIM: The aim of the study was to validate a rapid method to detect and quantify colistin resistance genes (mcr-1 to mcr-5) by real-time polymerase chain reaction (RT-PCR) in diverse matrices. METHODS AND RESULTS: The detection limit of two newly designed SYBR Green real-time PCR assays for mcr-4 and mcr-5 and of previously published protocols for mcr-1 to mcr-3 was assessed using serial dilutions of reference strains. The assays could detect all five mcr genes with the lower limit of 102 copy numbers. Escherichia coli isolates (n = 1062) and environmental samples (n = 93) were tested for the presence of mcr genes. The assays enabled the detection of colistin resistance genes both in bacterial isolates and in complex environmental samples. CONCLUSIONS: This method represents a set of sensitive, rapid and effective assays for the screening of colistin resistance directly from the environment. SIGNIFICANCE AND IMPACT OF THE STUDY: Colistin is an antimicrobial commonly used in animals and has recently emerged as a last-resort treatment in humans. Plasmid-mediated mcr genes confer resistance to colistin and represent a major threat for public health since they can be easily disseminated through horizontal gene transfer. The rapid and sensitive detection of mcr genes is of utmost necessity.

Serological evidence of H9N2 avian influenza virus exposure among poultry workers from Fars province of Iran.

BACKGROUND: Since the 1990s, influenza A viruses of the H9N2 subtype have been causing infections in the poultry population around the globe. This influenza subtype is widely circulating in poultry and human cases of AI H9N2 have been sporadically reported in countries where this virus is endemic in domestic birds. The wide circulation of H9N2 viruses throughout Europe and Asia along with their ability to cause direct infection in mammals and humans, raises public health concerns. H9N2 AI was reported for the first time in Iran in 1998 and at present it is endemic in poultry. This study was carried out to evaluate the exposure to H9N2 AI viruses among poultry workers from the Fars province. METHODS: 100 poultry workers and 100 healthy individuals with no professional exposure to poultry took part in this study. Serum samples were tested for antibodies against two distinct H9N2 avian influenza viruses, which showed different phylogenetic clustering and important molecular differences, such as at the amino acid (aa) position 226 (Q/L) (H3 numbering), using haemagglutination inhibition (HI) and microneutralization (MN) assays. RESULTS: Results showed that 17 % of the poultry workers were positive for the A/chicken/Iran/10VIR/854-5/2008 virus in MN test and 12 % in HI test using the titer ≥40 as positive cut-off value. Only 2 % of the poultry workers were positive for the A/chicken/Iran/12VIR/9630/1998 virus. Seroprevalence of non exposed individuals for both H9N2 strains was below 3 % by both tests. Statistical analyses models showed that exposure to poultry significantly increases the risk of infection with H9N2 virus. CONCLUSIONS: The results have demonstrated that exposure to avian H9N2 viruses had occurred among poultry workers in the Fars province of Iran. Continuous surveillance programmes should be implemented to monitor the presence of avian influenza infections in humans and to evaluate their potential threat to poultry workers and public health.

Differentiation of pityriasis lichenoides chronica from guttate psoriasis by dermoscopy.

Clinical differentiation between pityriasis lichenoides chronica (PLC) and guttate psoriasis (GP) may sometimes be a difficult task, which often requires histological analysis to reach a definitive diagnosis. In this study, PLC and GP lesions were examined using dermoscopy, and the significance of specific dermoscopic findings was investigated in order to facilitate their differentiation and decrease the number of cases requiring biopsy. We found that the incidence of orange-yellowish structureless areas, focal dotted vessels and nondotted vessels was statistically significant in PLC, while the incidence of diffuse dotted vessels was statistically significant in GP.

Effect of genotype, gender and feed restriction on growth, meat quality and the occurrence of white striping and wooden breast in broiler chickens.

Due to their importance for the control of meat quality in broiler chickens, the present study aimed at identifying the factors associated with the occurrence of myopathies and characterizing the meat properties when affected by myopathies. To this aim, a total of 768 broiler chickens were reared until slaughter (46 d) to evaluate the effect of genotype, gender, and feeding regime (ad libitum vs. restricted rate, 80% from 13 to 21 d of age) on performance and meat quality. Standard broilers were heavier (3,270 vs. 3,139 g; P<0.001) and showed lower feed conversion (1.56 vs. 1.61; P<0.001) than the high-yield broilers. Males showed higher final live weight (3,492 vs. 2,845 g) and lower feed conversion (1.54 vs. 1.63) than females (P<0.001). Feed restriction decreased final live weight (3,194 vs. 3,142 g; P<0.01) and feed conversion (1.60 vs. 1.57; P<0.01) compared to ad libitum feeding. At gross examination, feed restriction tended to increase white-striped breasts (69.5 vs. 79.5%; P<0.10), whereas females showed less wooden breasts than males (8.0 vs. 16.3%; P<0.05). White-striped fillets had higher pHu (5.87 vs. 5.83), and lower a* (-0.81 vs. -0.59) and b* color indexes (13.7 vs. 14.5) (P<0.05), whereas wooden breast fillets exhibited higher cooking losses (25.6 vs. 22.1%) and AK-shear force (4.23 vs. 2.84 kg/g) compared with normal fillets (P<0.001). At histological examination, 3.1% of pectoralis major were normal, 26.6% mildly degenerated, 45.3% moderately degenerated, and 25.0% severely degenerated. In conclusion, genotype had a moderate effect on growth without modifying myopathy occurrence. In contrast, gender and feed restriction affected performance, meat quality, and breast abnormalities.

Antimicrobial resistance and class 1 and 2 integrons in Escherichia coli from meat turkeys in Northern Italy.

This study is aimed at determining the antimicrobial resistance (AMR) and the presence of class 1 and 2 integrons in 48 avian pathogenic Escherichia coli (APEC) strains isolated from meat turkeys during three sequential production cycles. Thirty avian faecal E. coli (AFEC) strains from the first cycle were also analysed. Strains were tested for AMR against 25 antimicrobials by disk diffusion test and were screened for the presence of integrons and associated gene cassettes by polymerase chain reaction followed by sequencing. Genetic relatedness of isolates was established by pulsed-field gel electrophoresis. High levels of resistance were detected to tetracyclines, penicillins and sulphonamides in APEC and AFEC. Resistance to aminoglycosides, fluoroquinolones, cephalosporins and phenicols was variable, based on the antimicrobial drug and the isolate (APEC vs. AFEC). Full susceptibility to colistin was detected. Multidrug resistance of up to seven antimicrobial classes was exhibited by APEC (93.8%) and AFEC (100%). Nearly 44% of strains tested positive for class 1 and/or class 2 integrons containing the dfrA, aadA and sat2 genes, alone or in combination, coding for streptomycin/spectinomycin, trimethoprim and streptothricin resistance, respectively. The estX and orfF genes of unknown function were also detected. A significant association was found between the presence of integrons and the resistance to aminoglycosides and potentiated sulphonamides. The results of this study showed that AMR, multidrug resistance and class 1 and 2 integrons are widespread among pathogenic and commensal E. coli from Italian turkeys. More attention should be addressed to limit the use of antimicrobials in turkeys and the AMR of turkey E. coli.

Fluoroquinolone resistance and molecular characterization of gyrA and parC quinolone resistance-determining regions in Escherichia coli isolated from poultry.

Escherichia coli are a common inhabitant of the gastrointestinal tract of mammals and birds; nevertheless, they may be associated with a variety of severe and invasive infections. Whereas fluoroquinolones (FQ) have been banned in the United States for use in poultry production, the use of these antimicrobials in poultry husbandry is still possible in the European Union, although with some restrictions. The aim of this study was to investigate the FQ resistance of 235 E. coli isolates recovered from chickens and turkeys. Minimum inhibitory concentrations were determined by a microdilution method, whereas mutations in the quinolone resistance-determining regions of the target genes, gyrA and parC, were detected by a PCR-based method. High resistance rates (>60%) were observed for nalidixic acid, flumequine, and difloxacin, whereas resistance to ciprofloxacin, danofloxacin, enrofloxacin, marbofloxacin, and sarafloxacin was less frequently reported (<40%). Sixty-four isolates (27.2%) showed full susceptibility toward the tested FQ, but 57 isolates (24.2%) were resistant to all tested FQ. The remaining 114 E. coli isolates (48.5%) were grouped in 5 different resistance patterns. Isolates resistant only to flumequine or nalidixic acid or both possessed 1 gyrA mutation, whereas isolates with further resistance to enrofloxacin, difloxacin, danofloxacin, and sarafloxacin had in addition 1 or 2 parC substitutions. Two gyrA mutations coupled with 1 substitution in parC were detected in isolates resistant to all tested FQ. The number of mutations and their correlation with the in vitro activity of FQ reflected the currently accepted model, according to which a single gyrA substitution is associated with resistance or decreased susceptibility to older quinolones, whereas further gyrA or parC substitutions are needed for a higher level of resistance.

Enrofloxacin against Escherichia coli in turkeys: which treatment scheme is effective?

The efficacy of enrofloxacin (ENRO) was evaluated against multidrug-resistant avian pathogenic Escherichia coli correlating the minimum inhibitory concentrations (MIC) of 235 E. coli field strains with its pharmacokinetics (PK) in 50 healthy turkeys (5 groups) with a PK/pharmacodynamic approach. The treatments were as follows: a) single oral gavage and b) single subcutaneous (SC) treatment at the recommended dose of 10 mg/kg; c) single oral gavage, d) 5 d of 10-h pulsed water medication, and e) 5 d of 24-h continuous water medication at the doubled dose of 20 mg/kg. Blood samples were collected at established times over 24 h. Plasma was analyzed using a liquid chromatography tandem mass spectrometry method that was validated in house. A monocompartmental and a noncompartmental model were applied to the data to obtain the PK results. After gavage administration, the mean maximum concentration Cmax/MIC50 and area under the curve AUC0-24/MIC50 ratios were, respectively, 3.07 ± 0.62 and 7.01 ± 1.03 and 25.48 ± 3.04 and 57.2 ± 3.73 for the 10 and 20 mg/kg doses, respectively. After SC administration of 10 mg/kg, Cmax/MIC50 and AUC0-24/MIC50 ratios were 3.45 ± 0.75 and 33.96 ± 7.46, respectively. After the administration of 10-h pulsed or 24-h continuous medicated water at 20 mg/kg, lower values of Cmax/MIC50 (10-h pulsed: 3.45 ± 0.7; 24-h continuous: 3.05 ± 0.48) and AUC0-24/MIC50 (10-h pulsed: 42.42 ± 6.17; 24-h continuous: 53.32 ± 5.55) were obtained. Based on these results, the European Union-recommended dosage of 10 mg/kg seems ineffective to achieve adequate drug plasma concentrations and even the 20 mg/kg by 10 h pulsed or continuous medicated water administration did not reach completely efficacious concentrations in plasma against colibacillosis. Although the results obtained were not completely encouraging, the medicated water should preferably be provided continuously. To conclude about the efficacy of ENRO treatment against colibacillosis, target tissue concentration should be extensively considered.

A systematic review on the role of biosecurity to prevent or control colibacillosis in broiler production.

This systematic review aimed at investigating the role that biosecurity can have in preventing or controlling colibacillosis in broiler production. Primary studies with natural or experimental exposure to avian pathogenic Escherichia coli, evaluating any biosecurity measure to prevent or control colibacillosis in broiler chickens with at least one of the following outcomes: feed conversion ratio (FCR), condemnations at slaughter, and mortality due to colibacillosis, were included. A systematic search was carried out in 4 databases according to the Cochrane handbook and reported following the PRISMA 2020 directions. Studies (n = 3,886) were screened in a 2-phase process and data matching the inclusion criteria were extracted. Risk of bias assessment was performed. Four studies reporting biosecurity measures to prevent or control colibacillosis in broiler production were included. In all studies, only disinfection during either the pre-hatching period (n = 3) or the post-hatching period (n = 1) was evaluated as biosecurity measure in broiler production, as well as its effect on FCR (n = 2) and mortality (n = 4) due to colibacillosis. No studies with effects on condemnations at slaughter were found. Due to the heterogeneity of studies in regard to interventions and outcomes, meta-analysis was not carried out. The limited findings of this systematic review do not provide a comprehensive evidence to statistically evaluate the efficacy of biosecurity to prevent or control colibacillosis in broiler production. The scarcity of evidence found suggests that further and deeper investigations on the topic are needed, considering the variety of interventions related to biosecurity.

Pharmacokinetic/pharmacodynamic evaluation of the efficacy of flumequine in treating colibacillosis in turkeys.

Flumequine (FLU) is used in the treatment of systemic bacterial infections in poultry, including colibacillosis, which is a common disease in turkeys. The pharmacokinetic (PK) behavior of FLU administered to 32 healthy turkeys as an oral bolus via gavage or as 10-h pulsed administration in drinking water were compared, using the authorized dose of 15 mg/kg and the double dose of 30 mg/kg. The minimum inhibitory concentrations (MIC) of 235 Escherichia coli field strains isolated from poultry were determined for pharmacodynamics (PD) to develop a PK/PD model. Blood samples were collected at established times over 24 h, and the obtained plasma was analyzed using a liquid chromatography tandem mass spectrometry method that was validated in-house. A monocompartmental model and a noncompartmental model were applied to the data to obtain the PK results. For both types of administration and both dosages, the ratios of the maximum concentration (Cmax)/MIC50 and the area under the plasma concentration-time curve (AUC)/MIC50 achieved were considerably lower than the fluoroquinolone breakpoints usually adopted for efficacy. The Cmax/MIC50 and AUC0-24/MIC50 ratios were, respectively, 0.67 ± 0.09 and 4.76 ± 0.48 and 1.18 ± 0.35 and 7.05 ± 2.40 for the 15 and 30 mg/kg bolus doses, respectively. After 10-h pulsed administration of 15 mg/kg, values of Cmax/MIC50, 0.19 ± 0.02 on d 1 and 0.30 ± 0.08 on d 5 of therapy were obtained, the AUC/MIC50 ratios were 2.09 ± 0.29 and 3.22 ± 0.93 on d 1 and 5, respectively. Higher values were obtained with the doubled dose of 30 mg/kg: the Cmax/MIC50 ratios were 0.49 ± 0.11 on d 1 and 0.69 ± 0.18 on d 5; the AUC/MIC50 ratios were 5.15 ± 1.15 and 6.57 ± 1.92 on d 1 and 5, respectively. Based on these results, FLU administration should be adopted when specific diagnostic findings indicate its efficacy, and revising the dosage scheme to comply with the prudent and responsible use of antimicrobials in veterinary medicine is advisable.

Molecular characterization and genotypic antimicrobial resistance analysis of Campylobacter jejuni and Campylobacter coli isolated from broiler flocks in northern Italy.

Genetic variability and genotypic antimicrobial resistance (AMR) of Campylobacter jejuni and Campylobacter coli from commercial broiler farms were investigated in this study. Campylobacter isolates were genetically characterized by random amplified polymorphic DNA (RAPD)-polymerase chain reaction (PCR) and flaA-SVR and flaB-SVR sequence-based typing. Eight RAPD types were identified in C. jejuni and three in C. coli, while 16 fla profiles were detected among all isolates. Further, 13 flaA-SVR and 13 flaB-SVR alleles were identified. Both typing methods detected a high level of genetic diversity, but fla-SVR typing showed a higher discriminatory power. Indeed, Simpson's index of fla typing (D=0.920) was higher than that of RAPD typing (D=0.814). Moreover, the association of flaA-SVR and flaB-SVR sequence analysis showed a higher discriminatory power compared with the sequence analysis of single loci. Isolates were also analysed by the mismatch amplification mutation assay PCR test and the detection of cmeB gene to determine the occurrence of genetic determinants of AMR to macrolides and fluoroquinolones and multidrug resistance. The A2074C and A2075G mutations in the 23S rRNA gene, the C257T mutation in the gyrA gene, and the cmeB gene were higher in C. coli (19.0%, 67.0%, 100.0% and 100.0%, respectively) than in C. jejuni (0.0%, 3.1%, 48.3% and 48.3%, respectively). This study showed a high degree of genetic diversity and a high prevalence of genetic determinants of macrolide resistance, fluoroquinolone resistance and multidrug resistance among C. jejuni and C. coli isolates from Italian commercial broiler farms.

Mid-IR fiber-optic reflectance spectroscopy for identifying the finish on wooden furniture.

Mid-IR fiber-optic reflectance spectroscopy (FORS) is a totally noninvasive infrared analytical technique allowing the investigation of artworks without the need for any sampling. The development and optimization of this analytical methodology can provide a tool that is capable of supporting conservators during the first steps of their interventions, yielding fast results and dramatically reducing the number of samples needed to identify the materials involved. Furthermore, since reflection IR spectra suffer from important spectral anomalies that complicate accurate spectral interpretation, it is important to characterize known reference materials and substrates in advance. This work aims to verify the possibility of investigating and identifying the most widely used wood finishes by means of fiber-optic (chalcogenide and metal halides) mid-infrared spectroscopy. Two historically widely employed wood finishes (beeswax, shellac) and two modern ones (a hydrogenated hydrocarbon resin and a microcrystalline wax) were investigated in an extended IR range (from 1000 to 6000 cm(-1)) with reflectance spectroscopy and with FORS. The broad spectral response of the MCT detector was exploited in order to include overtones and combination bands from the NIR spectral range in the investigation. The reflectance spectra were compared with those collected in transmission mode in order to highlight modifications to shapes and intensities, to assign absorptions, and finally to select "marker" bands indicating the presence of certain finishing materials, even when applied onto a substrate such as wood, which shows many absorptions in the mid-infrared region. After the characterization, the different products were applied to samples of aged pear wood and investigated with the same techniques in order to check the ability of mid-IR FORS to reveal the presence and composition of the product on the wooden substrate.

Efficacy of locally delivered statins as an adjunct to scaling and root planning in the treatment of periodontitis: a systematic review and meta-analysis.

OBJECTIVE: The aim of this systematic review is to assess the efficacy of locally delivered statins used in adjunct to scaling and root planing (SRP), compared with SRP alone. MATERIALS AND METHODS: An electronic and hand search was carried out up to April 2020. Only randomized controlled trials (RCTs) were included. Clinical attachment level gain (CALgain) and probing depth reduction (PDred), modified sulcular bleeding index reduction (mSBIred), and intrabony defect reduction (IBDred) were the investigated outcomes. Meta-analysis was performed, and the power of the meta-analytic findings was determined by trial sequential analysis (TSA). Studies were also sub-grouped based on the type of statin used. Statistical heterogeneity and publication bias were assessed. RESULTS: Twenty RCTs were included (1212 patients, 1289 defects). An overall statistically significant effect size in favor of statins for CALgain and PDred was found. As opposed to atorvastatin and rosuvastatin, simvastatin did not reach statistical significance for these outcomes, as shown by the sub-group analysis. CONCLUSIONS: Within the limits of the available studies, the local administration of statins (in particular, atorvastatin and rosuvastatin) in adjunct to SRP may result in additional significant improvement in terms of CALgain and PDred compared with SRP alone. The high heterogeneity of data and the high risk of bias found, however, impose caution. No approved preparations, moreover, exist, and further well-designed RCTs from independent research centers are needed to confirm the beneficial effects of the different statins and their mutual differences in the non-surgical periodontal treatment.

Effect of feed restriction timing on live performance, breast myopathy occurrence, and muscle fiber degeneration in 2 broiler chicken genetic lines.

During recent years, research on meat quality in poultry has aimed to evaluate the presence and consequences of breast myopathies as well as the factors which can affect their occurrence by modifying the growth rate. A total of 900 broiler chickens were reared until slaughter (48 D) to evaluate the effect of 2 genetic lines (A vs. B) and feeding plans (ad libitum [AL], early restricted [ER], from 13 to 23 D of age, and late restricted [LR], from 27 to 37 D of age; restriction rate: 80%) on performance, meat quality, and breast muscle myopathies. Calsequestrin and vascular endothelial growth factor (VEGF) expressions, and muscle fiber degeneration (MFD) were recorded at 22, 36, and 48 D. Chickens in the AL treatment had greater final live (P < 0.01) and carcass weights and proportion of pectoralis major muscle (P = 0.04) compared to chickens in the LR treatment, whereas chickens in the ER treatment had intermediate final live (3,454 g) and carcass weights, and proportion of pectoralis major muscle (25.6%). Chickens of line A were heavier than chickens of line B (P < 0.001), and had a greater feed conversion rate. Chickens of line A also had a greater dressing out percentage (P < 0.001), but a lower proportion of pectoralis major muscle (P = 0.04), as well as a greater meat pH (P < 0.001), meat cooking losses (P < 0.01), and shear force of the pectoralis major muscle (P = 0.03). Calsequestrin and VEGF mRNA were significantly lower in ER and LR chickens compared to AL chickens after feed restriction and during refeeding (P < 0.05). MFD scores increased with chicken age (P < 0.001) and differed between genetic lines (P < 0.001). Neither feeding plan nor genetic line affected the occurrence of white striping or wooden breast condition.

Precocious puberty in Sanfilippo IIIA disease: diagnosis and follow-up of two new cases.

To date the association between mucopolysaccharidosis (MPS) IIIA and precocious puberty has been found in only three polish patients. We observed two children affected by MPS IIIA with central precocious puberty (CPP) both treated with GnRH agonists. The occurrence of CPP in both patients with MPS IIIA suggests that it is necessary to look for an association between the two conditions. The follow-up of our two patients leads us to believe also that GnRH agonist treatment can have a beneficial effect on final height and probably on the improvement of behavioural problems.

A Comparison between Piezoelectric Devices and Conventional Rotary Instruments in Bone Harvesting in Patients with Lip and Palate Cleft: A Retrospective Study with Clinical, Radiographical, and Histological Evaluation.

INTRODUCTION: Orofacial clefts are congenital malformations characterized by an incomplete shaping of structures that separate the nasal from the oral cavity and can affect the right, left, or both sides. The aim of the present study is to assess, with clinical, radiographical, and histological evaluations, the efficacy of piezoelectric devices compared to traditional rotating instruments in the bone harvesting in patients with history of cleft. MATERIALS AND METHODS: We have conducted a retrospective analysis on 20 patients with a history of orofacial clefts that were operated on from February 2014 to June 2017. The patients were divided into two groups: Group R in which bone graft was harvested using a burr and Group P in which the bone graft was obtained by a piezoelectric device. After a healing period of 8 months from the grafting procedure, clinical and radiographic evaluations were performed. RESULTS AND DISCUSSION: The use of the piezoelectric devices in bone harvesting allows a slight improvement in the final volume. This supports a faster integration into the receiving site. CONCLUSIONS: The use of piezoelectric device in patients with history of orofacial cleft that needed bone graft represents a method to be taken into consideration because it has interesting advantages.

Corrigendum to "A Comparison between Piezoelectric Devices and Conventional Rotary Instruments in Bone Harvesting in Patients with Lip and Palate Cleft: A Retrospective Study with Clinical, Radiographical, and Histological Evaluation".

[This corrects the article DOI: 10.1155/2018/2059464.].

Microbiological, chemical and physical quality of drinking water for commercial turkeys: a cross-sectional study.

Drinking water for poultry is not subject to particular microbiological, chemical and physical requirements, thereby representing a potential transmission route for pathogenic microorganisms and contaminants and/or becoming unsuitable for water-administered medications. This study assessed the microbiological, chemical and physical drinking water quality of 28 turkey farms in North-Eastern Italy: 14 supplied with tap water (TW) and 14 with well water (WW). Water salinity, hardness, pH, ammonia, sulphate, phosphate, nitrate, chromium, copper and iron levels were also assessed. Moreover, total bacterial count at 22°C, presence and enumeration of Enterococcus spp. and E. coli, presence of Salmonella spp. and Campylobacter spp. were quantified. A water sample was collected in winter and in summer at 3 sampling sites: the water source (A), the beginning (B) and the end (C) of the nipple line (168 samples in total). Chemical and physical quality of both TW and WW sources was mostly within the limits of TW for humans. However, high levels of hardness and iron were evidenced in both sources. In WW vs. TW, sulphate and salinity levels were significantly higher, whilst pH and nitrate levels were significantly lower. At site A, microbiological quality of WW and TW was mostly within the limit of TW for humans. However, both sources had a significantly lower microbiological quality at sites B and C. Salmonella enterica subsp. enterica serotype Kentucky was isolated only twice from WW. Campylobacter spp. were rarely isolated (3.6% of farms); however, Campylobacter spp. farm-level prevalence by real-time PCR was up to 43% for both water sources. Winter posed at higher risk than summer for Campylobacter spp. presence in water, whereas no significant associations were found with water source, site, recirculation system, and turkey age. Low salinity and high hardness were significant risk factors for C. coli and C. jejuni presence, respectively. These results show the need of improving sanitization of drinking water pipelines for commercial turkeys.

Effect of age on the occurrence of muscle fiber degeneration associated with myopathies in broiler chickens submitted to feed restriction.

To evaluate muscle fiber degeneration (MFD) associated with white striping and wooden breast, pectoralis major of 192 broilers differing for genotype (standard vs. high breast yield), gender, and feeding regime (ad libitum vs. restricted rate 80% from 13 to 21 d of age) were sampled at 14, 21, 28, 35, and 46 d of age for histological analyses by hematoxylin and eosin (H&E) staining to evaluate tissue morphology, Masson's trichrome to identify collagen presence, and Oil red and Nile blue for lipid presence. Microvessels (diameter ≤15 µm), nuclei positive to anti-cleaved lamin A and monoclonal proliferating cell nuclear antigen (PCNA) antisera were counted to assess apoptotic and regenerative processes, respectively. Significant differences were found according to feeding system, age, and their interactions. The frequency of chickens with MFD was higher with ad libitum than restricted feeding (75.0% vs. 62.5%; P = 0.01) and increased with age (18.8%, 28.1%, 75.1%, 96.9%, and 96.9% at 14, 21, 28, 35, and 46 d). However, at 14 d a similar frequency (18.8%) was found in all broilers; at 21 d, MFD occurred more in broilers fed ad libitum than in those under restriction (50.0% vs. 6.3%; P < 0.01); at 28 d differences were reduced (87.5% vs. 62.5%; P  = 0.10) to disappear by 35 (100% and 93.8%) and 46 d (96.9% and 96.9%). The number of microvessels decreased with age (20.7 to 9.46; P < 0.001) and the number of nuclei positive to the anti-cleaved lamin A antibody increased. At histology, MFD at 46 d corresponded to loss of typical cross striations, massive necrotic process, degenerating fibers surrounded by inflammatory cells, scattered fibers in an abundant collagen-rich connective tissue, numerous adipose cells; necrotic fibers showed a high percentage of apoptotic nuclei, and regenerating fibers appeared positive to anti-PCNA antibody. In conclusion, MFD soon occurred after 2 wk of growth and increased dramatically within 28 d. Early feed restriction reduced MFD as long as animals were restricted, but no residual effect was recorded after re-alimentation.

Survey of Campylobacter spp. in owned and unowned dogs and cats in Northern Italy.

Campylobacteriosis is among the most common bacterial causes of human gastroenteritis worldwide and pet ownership has been identified as a risk factor for Campylobacter infection in humans. Since canine and feline prevalence data are scarce in Italy, the present study was carried out to assess the prevalence, species distribution and risk factors for Campylobacter infection in dogs and cats under different husbandry conditions. Rectal swabs were collected from 171 dogs (household pets, n = 100; shelter-housed dogs, n = 50; dogs from breeding kennels, n = 21) and 102 cats (household pets, n = 52; shelter-housed cats, n = 21; free-roaming cats n = 29) in Northern Italy. Campylobacter was isolated from 17% (n = 29) of dogs and 14.7% (n = 15) of cats. C. jejuni was the most common isolate in both species (Campylobacter spp.-positive dogs, 55.2%; Campylobacter spp.-positive cats, 53.3%), followed by C. upsaliensis (Campylobacter spp.-positive dogs, 27.6%; Campylobacter spp.-positive cats, 40%). Other Campylobacter species were rarely detected, but included C. hyointestinalis subsp. hyointestinalis, C. lari and C. coli in dogs and C. coli and C. helveticus in cats. Among considered variables (sex, age, origin, diarrhoea, season of sampling), origin was identified as a risk factor for dogs, with shelter-housed dogs at higher risk than household dogs (odds ratio, 2.84; 95% CI 1.17, 6.92; P = 0.021). The results of this study, particularly the high prevalence of C. jejuni in Campylobacter-positive animals, demonstrated that household and stray dogs and cats in Northern Italy might pose a zoonotic risk for humans. Moreover, biosecurity measures should be improved in dog shelters.