Hazard Identification Related to the Presence of Vibrio spp., Biogenic Amines, and Indole-Producing Bacteria in a Non-Filter Feeding Marine Gastropod (Tritia mutabilis) Commercialized on the Italian Market

open4noTritia mutabilis is a carrion-feeder edible marine gastropod with an open circulatory sys- tem. Therefore, biological, and chemical contaminants associated with the feed can reach all body tissues. The aim of the present study was to investigate the possible association of these char- acteristics with some food safety hazards. Vibrio spp. load, and the prevalence of pathogenic V. parahaemolyticus, V. vulnificus, and V. cholerae, were investigated. Moreover, biogenic amines (BAs) and indole-producing bacteria (IPB), markers of seafood decomposition, were quantified for the first time in an edible carrion-feeder. Overall, 49 batches were analyzed (38 from retail, and 11 from primary production). The Vibrio spp. load resulted of 5.64 ±0.69 log10 CFU g−1 at retail, and 5.27 ±0.74 at harvest but all batches resulted negative for pathogenic Vibrio. Histamine, putrescine, cadaverine, and tyramine were detected both at harvest and at the retail level. Their sum (BAs Index) showed a mean value of 50.45 and 65.83 mg Kg−1 in batches at harvest and at retail, respec- tively. IPB were detected at harvest and upon refrigeration for three days (T1–T3). The mean load resulted in 2.52 ±0.85 log10 MPN g−1 at T0, 3.31 ±1.23 at T3 in batches immediately refrigerated, and 3.22 ±1.18 at T3 in batches previously immersed in clean seawater. Our results contribute to identifying food-borne hazards for T. mutabilis that may be related to the retention of biogenic amines and indole-producing bacteria due to carrion feedingopenSerratore, Patrizia; Bignami, Giorgia; Ostanello, Fabio; Lorito, LunaSerratore, Patrizia; Bignami, Giorgia; Ostanello, Fabio; Lorito, Lun

Improvement of environmental quality in intensive pig farming through an integrated bioactivation program for the control and prevention of swine mycoplasmal pneumonia

The decline of pig health status is the effect of a bad air quality inside breeding facilities related to the concentration of biogases derived from the action of faecal microbial flora on urine nitrogen. Bioactivation of the environment with bacterialenzymatic mixtures is able to reduce this chemical emanation. An experimental trial has been performed to evaluate the efficacy of the bioactivation treatment to prevent mycoplasmal pneumonia in finishing pigs. Treated and untreated groups, different treatment schemes and different floors have been compared. The ammonia concentration was checked by a chemical method. M. hyopneumoniae infection was evaluated through seroprevalence and by a quantitative lung-scoring system. For all slaughter-lots average carcass weight was recorded. The decrease of the environmental ammonia concentration resulted in lower lungscores and higher carcass weight

Detection and Characterization of Porcine Sapelovirus in Italian Pig Farms

Porcine sapelovirus (PSV) belongs to the genus Sapelovirus of the family Picornaviridae. PSV infects pigs asymptomatically, but it can also cause severe neurologic, enteric, and respiratory symptoms or reproductive failure. Sapelovirus infections have been reported worldwide in pigs. The objective of this study was to investigate the presence and the prevalence of PSV in Italian swine farms in animals of different ages to clarify the occurrence of the infection and the genetic characteristics of circulating strains. In the present study, 92 pools of fecal samples, collected from pigs across three farms, were analyzed by Reverse Transcriptase-polymerase Chain Reaction-PCR (RT-PCR). Fecal pools from young growers (63/64) were found positive for Sapelovirus in all farms while detection in sows (4/28) was observed in only one farm. Phylogenetic analyses of the 19 partial capsid protein nucleotide sequences (VP1) (6\u20137 each farm) enable the classification of the virus sequences into three distinct clades and highlighted the high heterogeneity within one farm. The whole genome sequence obtained from one strain showed the highest correlation with the Italian strain detected in 2015. The study adds novel information about the circulation and heterogeneity of PSV strains in Italy and considering the movement of pigs across Europe would also be informative for other countries

Detection of hepatitis E virus RNA in rats caught in pig farms from Northern Italy

Hepatitis E virus (HEV) strains belonging to the Orthohepevirus genus are divided into four species (A\u2013D). HEV strains included in the Orthohepevirus A species infect humans and several other mammals. Among them, the HEV\u20103 and HEV\u20104 genotypes are zoonotic and infect both humans and animals, of which, pigs and wild boar are the main reservoirs. Viruses belonging to the Orthohepevirus C species (HEV\u2010C) have been considered to infect rats of different species and carnivores. Recently, two studies reported the detection of HEV\u2010C1 (rat HEV) RNA in immunocompromised and immunocompetent patients, suggesting a possible transmission of rat HEV to humans. The role of rats and mice as reservoir of HEV and the potential zoonotic transmission is still poorly known and deserves further investigation. To this purpose, in this study, the presence of HEV RNA was investigated in the intestinal contents and liver samples from 47 Black rats (Rattus rattus) and 21 House mice (Mus musculus) captured in four pig farms in Northern Italy. The presence of both Orthohepevirus A and C was investigated by the real\u2010rime RT\u2010PCR specific for HEV\u20101 to HEV\u20104 genotypes of Orthohepevirus A species and by a broad spectrum hemi\u2010nested RT\u2010PCR capable of detecting different HEV species including rat HEV. The intestinal content from two Black rats resulted positive for HEV\u2010C1 RNA and for HEV\u20103 RNA, respectively. None of the House mice was HEV RNA positive. Sequence analyses confirmed the detection of HEV\u2010C1, genotype G1 and HEV\u20103 subtype e. The viral strain HEV\u20103e detected in the rat was identical to swine HEV strains detected in the same farm. Liver samples were negative for the detection of either rat HEV or HEV\u20103

Correlation of Anti-Salmonella Antibodies Between Serum and Saliva Samples Collected From Finisher Pigs

Saliva samples obtained by using absorptive devices, can provide an alternative diagnostic matrix to serum for monitoring disease status in pigs. The aim of this study was to investigate the correlation of anti-Salmonella antibodies between serum and saliva samples collected from pigs. Twenty individual paired serum and saliva samples were collected from a single farm. Anti-Salmonella IgG was detected in individual serum samples using a commercial Salmonella ELISA kit, validated for sera. The same kit was used with a protocol modified by extending incubation time and increasing temperature to test individual saliva samples. Anti-Salmonella IgG antibodies in pig saliva were always detected at a lower level than in the matching serum samples. A correlation (rho = 0.66; p = 0.002) and a moderate agreement (K > 0.62 p = 0.003) was found between individual Salmonella IgG in serum and saliva samples. Both correlation and the agreement levels are moderate. The size of this investigation was small, and further studies are necessary to further confirm these findings. The results of this work provide some evidence that saliva samples have the potential to be used for the diagnosis of Salmonella infection in pig farms

Enteropatia proliferativa da Lawsonia intracellularis nel suino

L\u2019enteropatia proliferativa del suino (proliferative enteropathy - PE), denominata anche ileite, \ue8 causata da Lawsonia intracellularis, un batterio intracellulare obbligato. La PE \ue8 una patologia a trasmissione oro-fecale che si manifesta soprattutto nella fase di magronaggio ed \ue8 responsabile di consistenti perdite economiche negli allevamenti intensivi. I danni economici sono causati dalla riduzione dell\u2019incremento ponderale e dell\u2019indice di conversione dell\u2019alimento e dall\u2019aumento della mortalit\ue0 e dei soggetti di scarto. La PE \ue8 endemica in numerosi Paesi con prevalenze di aziende ed animali infetti che, in Europa, superano il 90% e il 40%, rispettivamente. Nel suino, la patologia \ue8 caratterizzata da un ispessimento della mucosa intestinale dovuto alla proliferazione incontrollata delle cellule delle cripte intestinali accompagnata dall\u2019inibizione, ad opera di L. intracellularis, della maturazione e della differenziazione delle cellule caliciformi secretorie e delle cellule assorbenti. La conseguenza inevitabile \ue8 la riduzione dell\u2019assorbimento dei nutrienti e la perdita di aminoacidi e proteine nel lume intestinale, con conseguente diarrea. La PE si manifesta con due forme cliniche principali: 1) la forma acuta (enteropatia proliferativa emorragica - PHE) che si osserva principalmente in animali dai 4 ai 12 mesi d\u2019et\ue0, caratterizzata da una diarrea sanguinolenta e da elevata mortalit\ue0 (fino al 50%) e, 2) la forma cronica (adenomatosi intestinale - PIA) che colpisce suini dalle 6 alle 20 settimane d\u2019et\ue0, e il cui sintomo principale \ue8 rappresentato da una diarrea con feci pastose. Sono state descritte anche altre due forme cliniche a bassa incidenza: a) l\u2019enterite necrotica (NE), espressione di una forma di enteropatia proliferativa cronica complicata da infezioni secondarie che esita in un\u2019estesa necrosi coagulativa dell\u2019epitelio intestinale e, b) l\u2019ileite regionale (RI), risultante dalla guarigione delle lesioni dovute a NE e caratterizzata da deposizione di tessuto di granulazione e ispessimento della tonaca muscolare. La diagnosi indiretta, che viene generalmente eseguita utilizzando un test ELISA, consente di valutare l\u2019eventuale esposizione dei suini a L. intracellularis, mentre la diagnosi diretta (realizzata impiegando test biomolecolari qualitativi: PCR o quantitativi: qPCR, immunoistochimica - IHC) permette di valutare se l\u2019infezione \ue8 in atto. Analogamente a quanto accade per altre forme patologiche del suino, la semplice messa in evidenza di L. intracellularis nelle feci non rappresenta un criterio diagnostico valido nei confronti di PE. La corretta procedura diagnostica prevede la quantificazione del numero di microrganismi/grammo di feci e la messa in evidenza di L. intracellularis all\u2019interno delle lesioni intestinali. La profilassi e il controllo si basano sull\u2019applicazione di rigide misure di lavaggio e disinfezione che consentono di ridurre la contaminazione ambientale tra un ciclo produttivo e l\u2019altro e l\u2019applicazione di misure di biosicurezza interna. Particolare attenzione va riservata all\u2019alimentazione, che dovrebbe garantire l\u2019equilibrio della microflora intestinale tramite un corretto rapporto tra proteina altamente digeribile e frazione di fibra, con il supporto di integratori probiotici e prebiotici. In una logica di uso consapevole del farmaco, la somministrazione di massa di antibiotici durante la fase critica del magronaggio dovrebbe essere limitata ai soli gruppi con sintomatologia clinica, implementando invece la profilassi vaccinale.Porcine Proliferative Enteropathy (PE or ileitis) is an infectious enteric disease caused by the intracellular pathogen Lawsonia intracellularis (LI). PE is endemic in many countries and causes severe economic losses in swine production system worldwide due to reduction of daily weight gain, reduction of feed conversion ratio and increase of mortality and swine waste. In Europe, the prevalence of infected farms and infected animals is more than 90% and 40%, respectively. In PE, intestinal mucosa is thickened by uncontrolled proliferation of intestinal crypt cells while secretory cells and absorbent cells are decreased in number because LI prevents their maturation. Diarrhea is the consequence, due to reduced absorption and loss of amino acid and protein in intestinal lumen. Clinical forms are divided into acute (proliferative hemorrhagic enteropathy - PHE) and chronic form (porcine intestinal adenomatosis - PIA). Acute form affects animals from 4 to 12 weeks of age and is characterized by high mortality (>50%) and hemorrhagic diarrhea. Chronic form affects swine of 6-20 weeks of age and is characterized by pasty diarrhea. Based on morphological findings, two other forms are reported: necrotic enteritis (NE) and regional ileitis (RI). The first is a chronic form complicated by secondary infection that result in coagulative necrosis of intestinal epithelium. Healing of necrotic enteritis lesions results in both thickening of muscular layer of intestinal wall and granulation tissue deposition, both of which are typical findings of RI. Indirect diagnosis (e.g. ELISA) assess the exposure to L. intracellularis while direct diagnosis (PCR, qPCR, Immunohistochemistry -IHC) assess the current infection. Effective diagnosis is obtained comparing quantitation of microorganism/gram of feces with the detection of L. intracellularis within intestinal lesion. Prophylaxis and control of proliferative enteropathy are based on biosecurity measures combined with strict washing and disinfection measures to reduce environmental contamination. Proper nutrition management helps to ensure the balance of intestinal microflora by the use of highly digestible protein, by correct intake of fiber fraction and with probiotic and prebiotic supplements. To limit subclinical forms of disease, vaccination should replace antibiotic treatments which instead should be reserved only for symptomatic groups of pig

A screening sampling plan to detect Mycobacterium avium subspecies paratuberculosis-positive dairy herds

Abstract Mycobacterium avium ssp. paratuberculosis (MAP) is the etiological agent of paratuberculosis, a chronic contagious bacterial disease primarily affecting dairy cattle. Paratuberculosis represents a dual problem for the milk production chain: in addition to economic losses to affected herds, MAP may have zoonotic potential. Infected herds must be identified in order to implement programs designed to reduce the incidence of disease within and between herds and to prevent MAP from entering the food chain. The objective of this study was to evaluate the sensitivity and specificity of a screening sampling plan (SSP) to detect MAP-positive dairy herds by repetitive analysis of bulk tank milk (BTM) samples by ELISA and in-line milk filter (ILMF) samples by PCR. Samples from BTM and ILMF were collected twice from 569 dairy herds in southern Italy. Additionally, 12,016 individual milk samples were collected: 9,509 from 102 SSP-positive herds (SSP MAP-positive) and 2,507 from 21 randomly selected SSP-negative herds (SSP MAP-negative). There was a total of 126 SSP MAP-positive herds (i.e., 21.3% SSP MAP-positive herds; 95% confidence interval=18.0–24.9); the within-herd apparent prevalence (AP) ranged between 0.00 and 22.73% (mean 6.07%). A significant difference in within-herd AP was shown between SSP MAP-positive herds and SSP MAP-negative herds. A highly significant association was shown between the median AP herd status (>5%) and positivity to at least one ILMF or BTM sample. The SSP detected a minimum of 56.25% of low AP herds (AP ≤2.0%) up to a maximum of 100% of herds with a within-herd AP ≥8.0%. Overall, the SSP detected 85.57% of herds in which at least one individual milk sample was positive by ELISA. The proposed SSP was an inexpensive and useful tool to detect MAP-positive herds with a higher risk of infection diffusion and milk contamination. Although the SSP cannot be used for MAP-free certification of herds, it could be useful to prioritize appropriate control measures aimed at reducing the prevalence of infection in dairy herds and milk contamination

Evaluation of pre-slaughter losses of Italian heavy pigs

A retrospective observational study evaluated the risk factors for pre-slaughter losses (i.e. animal deaths occurring during transport and lairage) and their economic impact in Italian heavy pigs (about 160 kg body weight). Of the 3,344,730 pigs transported, 1,780 (0.053%) died before slaughter, with most losses occurring during transport (56.6%). The estimated economic impact was of \ue2\u82\uac 424,000. The percentage of batches with at least one animal lost pre-slaughter increased during summer (P<0.001). The proportion of pre-slaughter losses was higher when journey lasted above 90 min (P<0.001) and was correlated with transport duration (P<0.01). Losses were higher (P<0.01) in batches transported at low stocking densities (i.e., when heavier pigs were transported). Batches with lower slaughtering order (i.e., longer lairage time) had higher proportions of losses (P<0.001). Logistic regression analysis showed that the odds of a given batch to have at least one animal lost pre-slaughter were 1.32 times higher for batches slaughtered in summer, 1.54 times higher if journey durations exceeded 90 min, 1.25 times higher for batches with low slaughtering order, and not significantly influenced by stocking density during transport

Absence of Hepatitis E Virus (HEV) in Italian Lagomorph Species Sampled between 2019 and 2021

The zoonotic hepatitis E virus genotype 3 (HEV-3) causes most autochthonous human hepatitis E cases in Europe, which are due to the consumption of raw or undercooked food products of animal origin. Pigs and wild boars are considered the main reservoirs of this genotype, while rabbits are the reservoir of a distinct phylogenetic group named HEV-3ra, which is classified within the HEV-3 genotype but in a separate clade. Evidence for the zoonotic potential of HEV-3ra was suggested by its detection in immunocompromised patients in several European countries. HEV- 3ra infection was found in farmed and feral rabbit populations worldwide and its circulation was reported in a few European countries, including Italy. Furthermore, Italy is one of the major rabbit meat producers and consumers across Europe, but only a few studies investigated the presence of HEV in this reservoir. The aim of this study was to assess the presence of HEV in 328 Italian hares and 59 farmed rabbits collected in 3 Italian macro-areas (North, North-Central, and South-Central), between 2019 and 2021. For this purpose, liver samples were used to detect HEV RNA using broad- range real-time RT-PCR and nested RT-PCR. Using 28 liver transudates from hares, the ELISA test for anti-HEV IgG detection was also performed. Neither HEV RNA nor anti-HEV antibodies were detected. Further studies will be conducted to assess the HEV presence in Italian lagomorphs to establish the role of this host and the possible risk of transmission for workers with occupational exposure, to pet owners and via food

Occurrence of two Norovirus outbreaks in the same cafeteria in one week

In October 2017, two outbreaks of gastroenteritis (GE) occurred among patrons of a cafeteria in Italy in one week. Virological and bacteria investigations on stool samples, environment and food were conducted to identify the infectious agents and the possible source of infection. Forty-five cases occurred in the two outbreaks, including 13 laboratory-confirmed cases of norovirus GI. Nine staff members were interviewed, six were confirmed positive for NoV GI and 3 experienced GE symptoms. Bacteria faecal indicators and other bacteria pathogens were not detected in either environmental swab samples or food. A low level of NoV GII was detected in two environmental swab samples. The same GI.6 strain was identified in cases related to both outbreaks, suggesting a common source of infection. Since the two outbreaks occurred in one week, the NoV contamination could have persisted in the cafeteria. Furthermore, virological investigation revealed confirmed cases among food handlers who had worked at the cafeteria between and during the two outbreaks. Several studies highlighted the importance of excluding symptomatic food handlers to prevent contamination of foods and environment