Ecological and social factors influence interspecific pathogens occurrence among bees.

The interspecific transmission of pathogens can occur frequently in the environment. Among wild bees, the main spillover cases are caused by pathogens associated with Apis mellifera, whose colonies can act as reservoirs. Due to the limited availability of data in Italy, it is challenging to accurately assess the impact and implications of this phenomenon on the wild bee populations. In this study, a total of 3372 bees were sampled from 11 Italian regions within the BeeNet project, evaluating the prevalence and the abundance of the major honey bee pathogens (DWV, BQCV, ABPV, CBPV, KBV, Nosema ceranae, Ascosphaera apis, Crithidia mellificae, Lotmaria passim, Crithidia bombi). The 68.4% of samples were positive for at least one pathogen. DWV, BQCV, N. ceranae and CBPV showed the highest prevalence and abundance values, confirming them as the most prevalent pathogens spread in the environment. For these pathogens, Andrena, Bombus, Eucera and Seladonia showed the highest mean prevalence and abundance values. Generally, time trends showed a prevalence and abundance decrease from April to July. In order to predict the risk of infection among wild bees, statistical models were developed. A low influence of apiary density on pathogen occurrence was observed, while meteorological conditions and agricultural management showed a greater impact on pathogen persistence in the environment. Social and biological traits of wild bees also contributed to defining a higher risk of infection for bivoltine, communal, mining and oligolectic bees. Out of all the samples tested, 40.5% were co-infected with two or more pathogens. In some cases, individuals were simultaneously infected with up to five different pathogens. It is essential to increase knowledge about the transmission of pathogens among wild bees to understand dynamics, impact and effects on pollinator populations. Implementing concrete plans for the conservation of wild bee species is important to ensure the health of wild and human-managed bees within a One-Health perspective.

Effects of Virgin Coconut Oil-Enriched Diet on Immune and Antioxidant Enzymatic Activity, Fat and Vitellogenin Contents in Newly Emerged and Forager Bees (Apis mellifera L.) Reared in Cages.

Searching for artificial diets positively affecting the survival, immune and antioxidant systems of honey bees is one of main challenges occurring in beekeeping. Among nutrients, lipids play a significant role in insect nutrition as structural components in cell membranes, energy sources and reserves, and are involved in many physiological processes. In this context, the aim of this work was to investigate the effect of 0.5% and 1% coconut oil-enriched diet administration on newly emerged and forager bees survival rate, feed intake, immune system, antioxidant system and both fat and vitellogenin content. In newly emerged bees, supplementation with 1% coconut oil determined a decrease in feed consumption, an increase in survival rate from the 3rd to 14th day of feeding, a short-term decrease in phenoloxidase activity, an increase in body fat and no differences in vitellogenin content. Conversely, supplementation with 0.5% coconut oil determined an increase in survival rate from the 3rd to 15th day of feeding and an increase in fat content in the long term (i.e., 20 days). Regarding the forager bee diet, enrichment with 0.5% and 1% coconut oil only determined an increase in fat content. Therefore, supplementation with coconut oil in honey bee diets at low percentages (0.5 and 1%) determines fat gain. Further investigations to evaluate the use of such supplement foods to prevent the fat loss of weak families during winter are desirable.

Host-Parasitoid Relationship between Apis mellifera (Linnaeus, 1758) and Senotainia tricuspis (Meigen, 1838) (Diptera, Sarcophagidae): Fly Aggression Behavior and Infestation Rates of Senotainiosis.

Senotainia tricuspis (Meigen, 1838) is a Sarcophagid dipteran endoparasitoid of Apis mellifera L., and myiasis, caused by this fly, is reported in several European, North African and Middle Eastern countries. Nevertheless, very little knowledge concerning the aggression and parasitisation behavior of S. tricuspis toward A. mellifera is available in the scientific literature, and the temporal pattern of aggression remains unclear. The aim of this investigation was to describe the aggressive behavior of S. tricuspis and to provide data on pupation and adult emergence in order to identify further tools for the control of senotainiosis in beekeeping. Data were collected in an apiary in Pisa province (Tuscany, Italy), where observations of aggressive behavior were conducted indirectly by using a VHS camera and also directly by an observer. Four behavioral categories of the attack were described. A total of 55 aggressions, 21 beecatchers, 104 chases and 6 parasitization events were recorded with the camera. Slow-motion recording analyses of the parasitization episodes resulted in contact of at least 1/6 s between the parasitoid and the host. Through four days of direct observations, a total of 1633 aggression events were recorded. The daily temporal pattern of the number of aggressions showed two main peaks: one during the morning hours (i.e., from 10:00 to 11:00) and one in the afternoon (i.e., from 15:00 to 17:00). The morphometric data on the first-instars of S. tricuspis allowed us to hypothesize a penetration in the bee through its prothoracic spiracle as a modality of entrance in the host body. Third-instars successfully pupate when sinking in topsoil or clay soil, and adults emerge when left to a 4 °C overwintering period of six months. Furthermore, the high mortality rate of those larvae that did not sink and did not pupate successfully suggests that reaching a certain depth in the soil is a determining factor for larvae survival and that mulch and/or minimum soil tillage could prevent severe senotainiosis in apiaries.

Bombus terrestris terrestris (Linnaeus, 1758) and hybrids with the endemic Bombus xanthopus spotted on Capraia Island (Tuscan Archipelago, Italy): some conservation management implications.

Expansion of wild and managed allochthonous species leads to potential negative consequences for the endemic wildlife, such as resource competition, pathogens spread, hybridization and native species replacements. On Capraia Island, the last sighting of Bombus terrestris terrestris dates back to 1917. All subsequent surveys carried out on the island only reported the presence of B. xanthopus and B. pascuorum melleofacies with B. t. terrestris apparently no longer existing in the area. In 2021 B. t. terrestris was again detected on the island raising two main hypotheses: (i) B. t. terrestris has always been present with a low population density, such as not to be detected in previous investigations, or (ii) its presence is the result of a more recent recolonization. The recolonization event may be promoted by either intentional or unintentional introduction or it may be the result of a natural migration from the mainland or surrounding islands. Hybridization between B. t. terrestris × B. xanthopus on Capraia Island has been also ascertained by the detection of hybrid queens, workers and males. These new finding provides insight on the distribution range of B. t. terrestris in the Tuscan Archipelago and raise concern on the conservation of the endemic B. xanthopus population.

Effect of Ageing in the Mating Behaviour Sequence of Osmia cornuta Latr. (Hymenoptera: Megachilidae).

Osmia cornuta Latr. is largely managed worldwide for the pollination of orchard crops, playing a key role in the maintenance of healthy ecosystems and ensuring economic and social benefits for human society. The management techniques of this pollinator include the possibility of delaying emergence from cocoons after diapause, allowing for the pollination of later-blooming fruit crops. In this study, the mating behaviour of bees emerging at the natural time (Right Emergence Insects) and of late-emerged bees (Aged Emergence Insects) was described in order to test if a delay in emergence could affect the mating sequence of O. cornuta. Markov analysis of the mating behaviour revealed the occurrence of antenna motion episodes that were repeated in a stereotyped manner at regular intervals during the mating sequence of both Right Emergence Insects and in Aged Emergence Insects. Pouncing, rhythmic and continuous emission of sound, motion of antennae, stretching of the abdomen, short and long copulations, scratching, inactivity, and self-grooming were identified as the stereotyped behavioural units of a behavioural sequence. The occurrence of short copulations, the frequency of which increased with the age of bees, could lead to a failure in the reproduction of the mason bee.

Settlement of a stable wolf pack in a highly anthropic area of Pisan hills: Relationship with animal husbandry and hunting in a human-wolf coexistence perspective.

Anthropic areas play a pivot role for main wolf conservation challenges. Wolf presence in the higher Pisan hills has been well documented while wolf settlement in the lower Pisan hills is still uncertain. In this study, long-term information on wolf presence in a highly anthropic area of the lower Pisan hills was collected by using non-invasive monitoring techniques. Furthermore, both the relationship of this predator with human activity and the impact of hunting on wolf presence have been investigated. The results obtained indicate the presence of a stable and reproductive wolf pack composed by both Italian wolf and hybrids individuals in the municipalities of Crespina Lorenzana and Casciana Terme Lari. A high impact of wolf on livestock was recorded in this area since no prevention systems were adopted by farmers. Wolf appears not to have a negative impact on wild boar population. Similarly, wild boar drive hunting does not appear to affect the wolf pack presence in the area. Thereby wolf may play a key role as controller of wild population. Prevention strategies improvement becomes instrumental to promote wolf-human coexistence. Further investigation to monitor pack hybridization level and turnover and to assess the impact of packs on wild population and livestock in anthropic areas is desirable.

Taeniid cestodes in a wolf pack living in a highly anthropic hilly agro-ecosystem.

The Italian wolf population in human-modified landscapes has increased greatly in the last few decades. Anthropisation increases the risk of transmission of many zoonotic infections and in this context, control of taeniid cestode species needs to be addressed from a One Health perspective. Predator-prey interactions are at the root of taeniid cestode transmission, and the wolf plays a key role in the maintenance and transmission of taeniids. To date, all available data on the taeniids of wolves in Italy refer to populations living in a wild habitat. Between 2018 and 2019, we investigated taeniids in a wolf pack living in a highly anthropic hilly agro-ecosystem. Thirty-eight faecal samples were collected and analysed, 4 of which were also genetically characterised for individual wolves and belonged to three different animals. Samples collected were analysed microscopically and by molecular analysis in order to identify the taeniid species. Taeniid eggs were detected in 34.2% (13/38) of samples. Within samples positive to taeniid eggs only Echinococcus granulosus s.s. and Taenia hydatigena were identified in 26.3% and 10.5% of the samples, respectively. On microscopic examination, Capillaria spp., Ancylostomatidae and Toxocara canis eggs, Crenosoma vulpis larvae, and coccidian oocysts were also found. The combination of low biodiversity of taeniid species with a high occurrence of E. granulosus s.s. recorded in this study could be the consequence of a deeper link occurring between wolves and livestock in human-modified landscapes than in wild settings.

TITLE: Cestodes Taeniidae dans une meute de loups vivant dans un agroécosystème vallonné hautement anthropique. ABSTRACT: La population de loups italiens dans les paysages modifiés par l'homme a considérablement augmenté au cours des dernières décennies. L'anthropisation augmente le risque de transmission de nombreuses infections zoonotiques et, dans ce contexte, le contrôle des espèces de cestodes Taeniidae doit être abordé dans une perspective One Health. Les interactions prédateurs-proies sont à la base de la transmission des Cestodes Taeniidae, et le loup joue un rôle clé dans le maintien et la transmission des Taeniidae. À ce jour, toutes les données disponibles sur les Taeniidae du loup en Italie se réfèrent aux populations vivant dans un habitat sauvage. Entre 2018 et 2019, les Taeniidae d'une meute de loups vivant dans un agro-écosystème vallonné hautement anthropique ont été étudiés. Trente-huit échantillons fécaux ont été collectés et analysés, dont quatre ont également été génétiquement caractérisés pour des loups individuels et appartenaient à trois animaux différents. Les échantillons prélevés ont été analysés au microscope et par analyse moléculaire afin d'identifier les espèces de Taeniidae. Des œufs de Taeniidae ont été détectés dans 34,2 % (13/38) des échantillons. Dans les échantillons positifs aux œufs de Taeniidae, seul Echinococcus granulosus s.s. et Taenia hydatigena ont été identifiés dans 26,3 % et 10,5 % des échantillons, respectivement. Lors de l'examen microscopique, des œufs de Capillaria spp., d'Ancylostomatidae et de Toxocara canis, des larves de Crenosoma vulpis et des oocystes de coccidie ont également été trouvés. La combinaison d'une faible biodiversité d'espèces de Taeniidae avec une forte occurrence d'E. granulosus s.s. rapportées dans cette étude pourrait être la conséquence d'un lien plus profond entre le loup et le bétail dans les paysages humains que dans les paysages sauvages.

Unexpected Echinococcus multilocularis infections in shepherd dogs and wolves in south-western Italian Alps: A new endemic area?

The European distribution of Echinococcus multilocularis has been reported to be expanding east and north, whereas its southern limits are deemed stable. During a study on Echinococcus granulosus s.l. infections in wolves and dogs in the Italian Maritime Alps, we unexpectedly detected the presence of E. multilocularis eggs in four fecal samples from at least two shepherd dogs, and in five wolf fecal samples. This finding, in an area about 130 km south of the southernmost E. multilocularis report in the Alps, may suggest a rapid expansion southward. While infections in foxes are currently being investigated, these data seem to indicate the potential for a new E. multilocularis endemic area. If this will be confirmed, the implementation of surveillance programs in wild and domestic canids and preventative measures will become a priority.