A systematic review on the occurrence of Salmonella in farmed Tenebrio molitor and Acheta domesticus or their derived products.

Insects represent a sustainable and protein-rich food source. This new supply chain requires the study and monitoring of pathogens' presence and impact, as for other farmed animals. Among pathogens, Salmonella is of interest due to the well-established possibility for insects to harbor it. Since Acheta domesticus (cricket) and Tenebrio molitor (mealworm) are the most sold and farmed insect species, the present systematic review aimed to collect, select, and evaluate, in the available scientific literature, studies investigating the occurrence of Salmonella in these species sampled. All available studies published in peer-reviewed journals in English, French, Italian, Portuguese, German, and Spanish were considered. No time limits were imposed. We searched PUBMED, EMBASE, WEB of Science Core Collection, and Food Science and Technology Abstracts. The first date searched was May 10th, 2022; an update of the search was conducted on May 5th, 2023. The data synthesis was presented in tables reporting the number of positives on the number of total analyzed samples with other relevant characteristics of the study. The quality assessment was carried out considering relevant aspects for sampling and the method of analysis for Salmonella detection. At the end of the screening process, 10 and nine studies conducted on crickets and mealworms, respectively, were included for data extraction. The S. serovar Wandsworth and S. serovar Stanley were isolated only in one sample of ready-to-eat crickets. A second study detected OTUs related to S. enterica in cricket and mealworm powders. No studies detected Salmonella in mealworms according to cultural methods. The limitations of the present review are that few studies were retrieved and that included studies had important limitations in terms of study design as sampling was mostly based on convenience and not on a sound statistical basis. The present systematic review underlines the need to obtain reliable data about Salmonella presence in insects considering the growing market and the scaling up of existing farms. This research was funded by the Italian Ministry of Health - Ricerca Corrente IZSVe 03/21. The review protocol was published on the Systematic Reviews for Animals and Food (SYREAF) Web site (https://syreaf.org/protocols/).

A systematic review of experimental studies on Salmonella persistence in insects.

The consumption of insects as food and feed has been recently suggested as a possible alternative to the rising global food need, thus it is crucial to monitor any potential food safety hazards in the insect supply chain. The aims of this systematic review were to collect, select, and evaluate studies investigating the persistence of Salmonella in insects. We searched PUBMED, EMBASE, WEB of Science Core Collection, and Food Science and Technology Abstracts. In total, 36 papers investigating the persistence of Salmonella in insects (both holometabolous and heterometabolous) were included after screening. Regarding complete metamorphosis insects, the longest Salmonella persistence was reported in Phormia regina, in which the pathogen persisted for 29 days at 5 °C. Similarly, Salmonella persisted in the feces of Alphitobius diaperinus for 28 days. The incomplete metamorphosis insect showing the longest Salmonella persistence (>10 months) was Blatella germanica. Periplaneta americana excreted Salmonella via feces for 44 days until all the insects were dead. The retrieved data on the persistence of Salmonella can be useful for further analysis by risk assessors and decision-makers involved in the safety of insect-based food, contributing to defining the sanitary requirements and risk mitigation measures along the supply chain. The review protocol is registered in PROSPERO database (CRD42022329213).

A multi-center validation study on the discrimination of Legionella pneumophila sg.1, Legionella pneumophila sg. 2-15 and Legionella non-pneumophila isolates from water by FT-IR spectroscopy.

This study developed and validated a method, based on the coupling of Fourier-transform infrared spectroscopy (FT-IR) and machine learning, for the automated serotyping of Legionella pneumophila serogroup 1, Legionella pneumophila serogroups 2-15 as well as their successful discrimination from Legionella non-pneumophila. As Legionella presents significant intra- and inter-species heterogeneities, careful data validation strategies were applied to minimize late-stage performance variations of the method across a large microbial population. A total of 244 isolates were analyzed. In details, the method was validated with a multi-centric approach with isolates from Italian thermal and drinking water (n = 82) as well as with samples from German, Italian, French, and British collections (n = 162). Specifically, robustness of the method was verified over the time-span of 1 year with multiple operators and two different FT-IR instruments located in Italy and Germany. Moreover, different production procedures for the solid culture medium (in-house or commercial) and different culture conditions (with and without 2.5% CO2) were tested. The method achieved an overall accuracy of 100, 98.5, and 93.9% on the Italian test set of Legionella, an independent batch of Legionella from multiple European culture collections, and an extra set of rare Legionella non-pneumophila, respectively.

Bombyx mori from a food safety perspective: A systematic review.

Bombyx mori (BM) is an economically important insect for silk production, and it is also farmed and used as food in different countries around the world. The present systematic review aims to assess the suitability of BM as an edible insect, retrieving data from scientific papers reporting microbiological, chemical, and allergenic hazards of silkworm consumed as either whole insects or insect derivatives. We considered all studies published in peer-reviewed journals in English, French, and Spanish languages. No time limits were imposed. We searched PUBMED, WEB of Science Core Collection, and EMBASE databases. The last literature search was carried out on May 5th, 2021. Data were collected in pre-defined tabular forms for the aforementioned hazards. In total, 65 records investigating the safety aspects were included after screening: 32 on microbiological hazards; 27 on chemical hazards; 16 on allergenic hazards. Concerning microbiological aspects, a high presence of Enterococcus in raw insects (5.00 % to 70.10 %) was reported through metagenomic analysis. Through non-metagenomic methods (classical and biomolecular microbiology techniques), Bacillus cereus and Pseudomonas fluorescens were the most commonly investigated and detected bacteria in the unprocessed insects, while B. cereus and Enterobacteriaceae were studied and reported in insect-based food. The foodborne pathogens Listeria monocytogenes and Salmonella spp. were never detected. Concerning toxicological aspects, three studies assessed the toxicity of BM powder in laboratory animals, but no negative effects were observed. Regarding heavy metal bioaccumulation in BM, evidence was reported for As, Cu, and, Zn. Allergic reactions following the ingestion of BM or derivative products are due to proteins that are widespread in arthropods. Furthermore, BM proteins can undergo possible cross-reactions with proteins of other insect species or crustaceans. However, heat treatments do not seem to reduce the allergenic potential of the silkworm proteins. The major limitation of the present review is that we could include only scientific literature published in Western languages, while the majority of relevant studies were conducted in Asian countries and part of them are published in Asian languages. In conclusion, scientific evidence regarding microbiological and chemical hazards of BM relevant for food safety is very limited. In the present work microbiological and chemical hazards relevant for food safety were identified in BM, however their presence do not impair the use as food but suggest the need for a risk assessment under specific conditions of production and use. Allergic reactions are possible in sensitive individuals following the ingestion of edible BM.

Authentication of Edible Insects' Powders by the Combination of DART-HRMS Signatures: The First Application of Ambient Mass Spectrometry to Screening of Novel Food.

This feasibility study reports the use of direct analysis in real-time high-resolution mass spectrometry (DART-HRMS) in profiling the powders from edible insects, as well as the potential for the identification of different insect species by classification modeling. The basis of this study is the revolution that has occurred in the field of analytical chemistry, with the improved capability of ambient mass spectrometry to authenticate food matrices. In this study, we applied DART-HRMS, coupled with mid-level data fusion and a learning method, to discriminate between Acheta domesticus (house cricket), Tenebrio molitor (yellow mealworm), Locusta migratoria (migratory locust), and Bombyx mori (silk moth). A distinct metabolic fingerprint was observed for each edible insect species, while the Bombyx mori fingerprint was characterized by highly abundant linolenic acid and quinic acid; palmitic and oleic acids are the statistically predominant fatty acids in black soldier fly (Hermetia illucens). Our chemometrics also revealed that the amino acid proline is a discriminant molecule in Tenebrio molitor, whereas palmitic and linoleic acids are the most informative molecular features of the house cricket (Acheta domesticus). Good separation between the four different insect species was achieved, and cross-validation gave 100% correct identification for all training samples. The performance of the random forest classifier was examined on a test set and produced excellent results, in terms of overall accuracy, sensitivity, and specificity. These results demonstrate the reliability of the DART-HRMS as a screening method in a future quality control scenario to detect complete substitution of insect powders.

Nutritional Composition of Bombyx mori Pupae: A Systematic Review.

As insects have started to enter the eating habits of Western countries, an increasing amount of literature regarding the mulberry silkworm (Bombyx mori) prospective application as food has been published. Despite this growing interest, there is currently no systematic review of silkworm nutritional composition available. In this paper, we performed a systematic review of the recent available literature on the nutrient composition of mulberry silkworm pupae. After screening the titles and abstracts of 14,008 studies retrieved from three scientific databases, data about nutrients was extracted from 29 selected papers, together with their related variables. This systematic review provides an overview of the variety of data reported in the literature and highlights that many elements contribute to hindering a sound comparison of the different nutritional values reported for silkworm pupae. The observed variability of the composition data reported could be due to differences in diet, strains, pretreatments, and origin of the silkworm analyzed. However, all these variables were not always available and should be reported in future studies to simplify the data comparison.

Assessing direct analysis in real-time mass spectrometry for the identification and serotyping of Legionella pneumophila.

AIMS: The efficacy of ambient mass spectrometry to identify and serotype Legionella pneumophila was assessed. To this aim, isolated waterborne colonies were submitted to a rapid extraction method and analysed by direct analysis in real-time mass spectrometry (DART-HRMS). METHODS AND RESULTS: The DART-HRMS profiles, coupled with partial least squares discriminant analysis (PLS-DA), were first evaluated for their ability to differentiate Legionella spp. from other bacteria. The resultant classification model achieved an accuracy of 98.1% on validation. Capitalising on these encouraging results, DART-HRMS profiling was explored as an alternative approach for the identification of L. pneumophila sg. 1, L. pneumophila sg. 2-15 and L. non-pneumophila; therefore, a different PLS-DA classifier was built. When tested on a validation set, this second classifier reached an overall accuracy of 95.93%. It identified the harmful L. pneumophila sg. 1 with an impressive specificity (100%) and slightly lower sensitivity (91.7%), and similar performances were reached in the classification of L. pneumophila sg. 2-15 and L. non-pneumophila. CONCLUSIONS: The results of this study show the DART-HMRS method has good accuracy, and it is an effective method for Legionella serogroup profiling. SIGNIFICANCE AND IMPACT OF THE STUDY: These preliminary findings could open a new avenue for the rapid identification and quick epidemiologic tracing of L. pneumophila, with a consequent improvement to risk assessment.

Prevalence of SARS-CoV-2 RNA on inanimate surfaces: a systematic review and meta-analysis.

Coronavirus disease (COVID-19) is a respiratory disease affecting many people and able to be transmitted through direct and perhaps indirect contact. Direct contact transmission, mediated by aerosols or droplets, is widely demonstrated, whereas indirect transmission is only supported by collateral evidence such as virus persistence on inanimate surfaces and data from other similar viruses. The present systematic review aims to estimate SARS-CoV-2 prevalence on inanimate surfaces, identifying risk levels according to surface characteristics. Data were obtained from studies in published papers collected from two databases (PubMed and Embase) with the last search on 1 September 2020. Included studies had to be papers in English, had to deal with coronavirus and had to consider inanimate surfaces in real settings. Studies were coded according to our assessment of the risk that the investigated surfaces could be contaminated by SARS-CoV-2. A meta-analysis and a metaregression were carried out to quantify virus RNA prevalence and to identify important factors driving differences among studies. Thirty-nine out of forty retrieved paper reported studies carried out in healthcare settings on the prevalence of virus RNA, five studies carry out also analyses through cell culture and six tested the viability of isolated viruses. Overall prevalences of SARS-CoV-2 RNA on high-, medium- and low-risk surfaces were 0.22 (CI95 [0.152-0.296]), 0.04 (CI95 [0.007-0.090]), and 0.00 (CI95 [0.00-0.019]), respectively. The duration surfaces were exposed to virus sources (patients) was the main factor explaining differences in prevalence.

Coagulase negative staphylococci from ovine bulk-tank milk: Effects of the exposure to sub-inhibitory concentrations of disinfectants for teat-dipping.

Teat-dipping is one of the most effective methods to prevent mammary infections in ruminants, including sub-clinical mastitis caused by coagulase-negative staphylococci (CoNS). Improper disinfectant application could expose microorganisms to sub-inhibitory concentrations leading to phenotypic variations. In this study, 12 chlorhexidine-digluconate (CHDG)-tolerant (of which 4 qac positive) and 12 benzalkonium chloride (BC)-tolerant (of which 7 qac-positive) CoNS isolates from ovine milk were exposed to sub-inhibitory concentrations of CHDG and BC, respectively. Changes in disinfectant susceptibility against BC and CHDG, antibiotic resistance against 12 antibiotics and biofilm production were then assessed for both groups. After CHDG stress, 67 % and 83 % of the CHDG-stressed isolates doubled their MICs for BC and CHDG, respectively and 2 qac-negative isolates showed a four-fold increase of their MBCs for CHDG. After BC stress, MICs for BC and CHDG doubled in 58 % and 83 % of the BC-stressed isolates, respectively, while one qac-positive isolate increased four-fold the MIC for BC. Cross-resistance to antibiotics was assessed by disc diffusion method. Some qac-positive isolates varied their resistance profile, while a blaZ-positive isolate showed a resistant phenotype against ampicillin only after the exposure to the disinfectant. As for qac-positive isolates, one CHDG-stressed and 2 BC-stressed increased their resistance to kanamycin and cefoxitin, respectively. The Congo Red Agar test was carried out to assess the in vitro slime production: all isolates were negative after stress. In conclusion, sub-inhibitory exposure to disinfectants may affect disinfectant and antibiotic susceptibility, the latter in particular for qac-positive isolates and those hosting unexpressed antibiotic resistance genes.

A systematic review of human coronaviruses survival on environmental surfaces.

The current pandemic caused by Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) has led people to implement preventive measures, including surface disinfection and use of alcohol-based hand gel, in order to avoid viral transmission via fomites. However, the role of surface transmission is still debated. The present systematic review aims to summarize all the evidence on surface survival of coronaviruses infecting humans. The analysis of 18 studies showed the longest coronavirus survival time is 28 days at room temperature (RT) on different surfaces: polymer banknotes, vinyl, steel, glass, and paper banknotes. Concerning SARS-CoV-2 human infection from contaminated surfaces, dangerous viral load on surfaces for up to 21 days was determined on polymer banknotes, steel, glass and paper banknotes. For viruses other than SARS-CoV-2, the longest period of survival was 14 days, recorded on glass. Environmental conditions can affect virus survival, and indeed, low temperatures and low humidity support prolonged survival of viruses on contaminated surfaces independently of surface type. Furthermore, it has been shown that exposure to sunlight significantly reduces the risk of surface transmission. Although studies are increasingly investigating the topic of coronavirus survival, it is difficult to compare them, given the methodology differences. For this reason, it is advisable to define a reference working protocol for virus survival trials, but, as an immediate measure, there is also a need for further investigations of coronavirus survival on surfaces.

Next generation sequencing study on RNA viruses of Vespa velutina and Apis mellifera sharing the same foraging area.

The predator Asian hornet (Vespa velutina) represents one of the major threats to honeybee survival. Viral spillover from bee to wasp has been supposed in several studies, and this work aims to identify and study the virome of both insect species living simultaneously in the same foraging area. Transcriptomic analysis was performed on V. velutina and Apis mellifera samples, and replicative form of detected viruses was carried out by strand-specific RT-PCR. Overall, 6 and 9 different viral types were reported in V. velutina and A. mellifera, respectively, and five of these viruses were recorded in both hosts. Varroa destructor virus-1 and Cripavirus NB-1/2011/HUN (now classified as Triato-like virus) were the most represented viruses detected in both hosts, also in replicative form. In this investigation, Triato-like virus, as well as Aphis gossypii virus and Nora virus, was detected for the first time in honeybees. Concerning V. velutina, we report for the first time the recently detected honeybee La Jolla virus. A general high homology rate between genomes of shared viruses between V. velutina and A. mellifera suggests the efficient transmission of the virus from bee to wasp. In conclusion, our findings highlight the presence of several known and newly reported RNA viruses infecting A. mellifera and V. velutina. This confirms the environment role as an important source of infection and indicates the possibility of spillover from prey to predator.

Detection of Pseudorabies Virus in Wild Boar Foetus.

Pseudorabies, or Aujeszky's disease, is a notifiable worldwide infection of domestic and feral swine that causes economic losses for the swine industry. In domestic pigs, the virus is responsible for nervous and/or respiratory symptoms; in pregnant sows, it is one of the major causes of stillbirth, mummification, embryonic death, and infertility (SMEDI). It is known that PRV infection in wild boar is associated with low pathogenicity and attenuated or absent symptomatology, but limited information is available about the ability of the virus to infect the foetuses of infected wild boar pregnant sows. Due to scarce information about the reproductive consequences, we investigate the possible intrauterine vertical transmission of the virus in wild boar pregnant sow living in a highly infected area. A number of 54 hunted wild boar were sampled during 2018-2019, and blood, genital and nasal swabs, placenta, and fetuses were collected for serological and molecular investigations. A seroprevalence of 74% (40/54) was detected, while 1/24 pregnant sow and 1/24 pooled foetuses tested positive by PCR (gene gB). This is the first evidence of viral detection in foetuses from seropositive pregnant wild boar. This finding suggests the possible pathogenetic role of PRV on pregnancy in wild boar and the existence of an additional transmission route.