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Selection bias can occur at two places during the review

Selection bias can occur at two places during the review process: (1) when decisions are made about where to obtain relevant literature and (2) when decisions are made about which studies to include in the review once the studies are identified. Two tools can be used to reduce selection bias. The first tool is an extensive search and the second tool is a predefined scope and relevance (eligibility) criteria. These tools could be used in any review to clarify to the end user the potential for selection bias in the review. Even if these tools (described below) are not employed, as might frequently be the case, authors could identify the source of studies for the end user. For example, authors might say the literature used was based on the author\’s encyclopedic knowledge of relevant studies, the literature was identified using a snowball approach i.e. gathered unsystematically based on interest, the literature was simply found by scanning the literature haphazardly for relevant studies, or author-based searches were used (Foster, 2003).
An example of a transparent description used in a narrative review is as follows: ‘A large proportion of this tie2 review article is based on the author\’s experience, opinions, and data which have only been published and presented in abstract form’ (Berent, 2011). Such a transparent description enables assessment of selection bias and acknowledges that the review is likely not reproducible.
Note that for this narrative integrative review about research synthesis in veterinary science, the literature were obtained in the following way: initially, we obtained a recent relevant paper from our own collection (Whittemore et?al., 2014) and identified the MESH terms used for that paper. One author then searched PubMed using the search strings ‘Review Literature as Topic’[MAJR] (3045 citations) and (‘Meta-Analysis as Topic’[MAJR] (3672 citations) and modified the search to reduce the number of papers. The final search string in PubMed was (‘Bias (Epidemiology)’[MeSH Terms]) AND (‘Meta-Analysis as Topic’[MAJR] OR ‘Review Literature as Topic’[MAJR])\’ and resulted in 568 citations.
The 568 papers were screened for opinion papers about research synthesis; papers solely about systematic reviews or meta-analysis were excluded. Twenty-two articles were considered potentially relevant and the full text of 18 articles were evaluated for relevance (Felson, 1992; Hutchison, 1993; Oxman and Guyatt, 1993; Oxman, 1994; Gilbody et?al., 1995; Slavin, 1995; Mosteller and Colditz, 1996; Joyce et?al., 1998; Raymond et?al., 1998; Williams, 1998; Evans and Kowanko, 2000; Ezzo et?al., 2001; Chalmers et?al., 2002; Foster, 2003; Thomas et?al., 2004; Nunn, 2008; Ioannidis and Mullard, 2014; Whittemore et?al., 2014). The remaining articles cited were found by using the references cited by other studies and our collection, which had been obtained over 10 years of haphazardly collecting articles related to the conduct of reviews and from writing our own publications.

Information bias in reviews
Just as in any primary research, incorrect information will result in bias in research synthesis (Elamin et?al., 2009; Tripepi et?al., 2010). For most narrative integrative reviews cardiac muscle is unclear how, or even if, the accuracy of the reported results from the primary studies is verified. In all reviews, steps should be taken to ensure that the inference drawn from the primary research is correct and end users should be made aware of the steps employed. Although the peer-review process is designed to assess the accuracy of the review results, it is beyond the scope of editors and peer reviewers to verify all statements made about the results of the primary research in the review. If the reviewer is particularly knowledgeable about the topic, he or she might identify some errors, but finding reviewers with encyclopedic knowledge of each review topic is very difficult. Two tools are available for reducing information bias in reviews, namely, duplicate data extraction and explicit data extraction.

In and bear meat has been illegally

In 1998 and 2005, bear meat has been illegally imported in personal baggage from Alaska and Canada to Germany and France, respectively. Since the Trichinella spp. larval burden in bear muscles can be very high, patients who acquire trichinellosis by the consumption of this meat may suffer from a more serious illness; it follows that outbreaks caused by bear meat are frequently described (Ancelle et al., 2005; Houzé et al., 2009; Hall et al., 2012). Certainly, the number of illegally imported meat items infected by Trichinella sp. is higher than the number of documented cases but it is likely that a percentage of these infected meat products are consumed well-cooked, thus preventing human infections.
Trichinella infected animals or meat which were legally or illegally introduced in a country were the source of 3443 Trichinella infections in humans in a 40-year period (1975–2014). Most of these infections (96.8%) have been linked to horsemeat consumption, whereas meat from pigs, wild boars and bears accounted only for 2.2%, 0.7% and 0.3% of cases, respectively.
The bush meat illegally imported from Africa to Europe can represent another way of importation of Trichinella sp. infected meat. In 2008, it was estimated that about five tons of bush meat per week were illegally imported in personal baggage from Africa to Europe through the Charles de Gaulle airport in Paris, France (Chaber et al., 2010). From 2008 to 2011, an average year weight of 5.5ton of meat of which 1.4% was bush meat, has been confiscated at Swiss international airports (Falk et al., 2013). Meat and meat derived products of pig origin illegally introduced by personal baggage from Asia to the European Union have been discovered in two German international airports from 2012 to 2013 (Beutlich et al., 2015). These three examples show the high risk for the introduction of Trichinella sp. infected meat which can be the source of human outbreaks thwarting the efforts made in the EU to control these zoonotic flap inhibitor in the food chain.
The importation of live animals such as polar bears caught in the wild to be kept in captivity in zoos does not represent a direct risk for the human health if the animal carcass is properly destroyed when the animal dies. If the animal carcass or some of its parts are improperly used to feed other animals or are released into the environment, Trichinella sp. larvae can be ingested by local animals establishing a new focus of these parasites. Fortunately, in the last decades, the collection of wild animals kept in captivity at zoos has been reduced and, today, most animals present in zoos are born in captivity (Gippoliti, 2012; Bilski et al., 2013). Today, the collection of live animals from the wild concerns mainly endangered species which are kept in captivity for their protection, reproduction and release into the wild (Lees and Wilcken, 2009; Marsden et al., 2013).
The migration of wild animals susceptible to Trichinella spp. infections can represent another way of the introduction (Széll et al., 2013) or increased prevalence (Pannwitz et al., 2010) of these zoonotic parasites into a country. The passive and/or active introduction of allopatric species which are susceptible hosts for Trichinella spp. can represent an important threat for the meat industry and food safety. The American mink carrier of T. pseudospiralis in the Danish island of Bornholm represents an emblematic example.
Integration of veterinary and public health efforts, i.e. the one health concept, is needed to monitor Trichinella sp. infection and to develop a comprehensive food safety program for these zoonotic nematodes (Bidaisee and Macpherson, 2014). Joining different authorities and expertise in a collaborative effort can be far more effective in controlling these parasites. The trackback of human cases is extremely important for veterinary services to carry out the investigations needed to identify sources of animal infection and to take the measures necessary to eliminate the infection. In turn, veterinary services should regularly inform public health agencies about data obtained from the surveillance and monitoring activities for meat-borne parasites in livestock and wildlife (Murrell, 2013).

The A galli sequences had a positive Fu

The A. galli sequences had a positive Fu’s Fs value of 0.625 (P=0.016) and Tajima’s D value of 1.842 (>0.05). Mismatch distribution analysis of the complete datasets revealed the presence of a multi-peak (Fig. 1), which suggests that there was no rapid expansion event that occurred in the South African’s A. galli population’s demographic history and a gene flow (Nm) value of −30.53 was observed. The large and negative Nm value in this study indicated less gene flow among the A. galli populations from the two provinces over time suggesting less movement of chickens between provinces. Höglund et al. (2012) observed a relatively high gene flow amongst Swedish A. galli isolates in buy Cycloheximide to our findings. The positive values from Tajima’s D test signify that A. galli might not have experienced population expansion in the past. The Tajima’s D and mismatch distribution analysis confirmed that A. galli from the two provinces were genetically similar and could not be considered as distinct populations. The high level of genetic similarity between the KwaZulu-Natal and Limpopo populations could therefore be due to other factors other than gene flow such as overlapping selection pressures and common founder effects between populations.
Four of the six haplotypes were shared between the two provinces (Fig. 2). Katakam et al. (2010) reported the presence of three A. galli haplotypes (I, II and III). Network analysis revealed that the six haplotypes observed in this study clustered separately from each other and were novel in comparison to those found in Genbank. Four (V, VI, VII and VIII) South African haplotypes were shared between isolates from Limpopo and KwaZulu-Natal provinces whilst haplotypes IV and IX were restricted to Limpopo province. Haplotype VIII had the highest frequencies of 14 A. galli isolates followed by haplotype V (n=13), VI buy Cycloheximide (n=6) and VII (n=5) whereas haplotype IV and IX consisted of only one individual each (Fig. 2).
This study presents the first attempt to genetically characterize A. galli parasites from village chicken populations of South Africa. The genetic similarity amongst the South African isolates could be useful in implementing helminths management and control strategies between South African provinces. The haplotypes observed in South Africa were different from those from Denmark which suggests that unique A. galli control strategies might be necessary rather than replicating control regimes from other areas. Further characterization of the worms is required particularly focusing on pathogenicity profiles.

Conflict of interest

Author contributions

Ethical standards

This study was funded jointly by the Agricultural Research Council-BTP and the NRF under the Zambia/South Africa bilateral Research Program. Ms Maltji received an NRF-DST Professional Development Program research fellowship and University of Pretoria postgraduate support bursary.

Besnoitia besnoiti—the cause of bovine besnoitiosis—is a cyst forming apicomplexan parasite closely related to Toxoplasma gondii and Neospora caninum (Ellis et al., 2000). In acutely infected cattle, bovine besnoitiosis is characterized by pyrexia, swollen lymphnodes, nasal and ocular discharge, salivation, stiff gait and—in severe cases—clinically apparent subcutaneous edema. In chronically infected cattle, the skin may become severely lichenified and alopecic. Bulls may become infertile due to orchitis (Bigalke and Prozesky, 2004; Kumi-Diaka et al., 1981). However, only a few cattle in affected herds develop typical clinical signs, while most animals remain subclinically infected (Alvarez-Garcia et al., 2013; Bigalke, 1968; Jacquiet et al., 2010). In the acute stage of the infection, tachyzoites multiply mainly in endothelial cells of blood vessels (McCully et al., 1966). Later on, characteristic tissue cysts establish, which are surrounded by hyaline material consisting of multiple layers of host cell-derived collagen fibrils, thus forming a secondary cyst wall (Dubey et al., 2013; Majzoub et al., 2010). Tissue cysts are located in the skin, the scleral conjunctivae, the mucosal membrane of the vestibulum vaginae and the upper respiratory tract (Basson et al., 1970; McCully et al., 1966). In addition, tissue cysts are regularly found in the aponeuroses of muscles and in fasciae (Basson et al., 1970; Gentile et al., 2012; McCully et al., 1966). Interestingly, internal organs are only rarely affected (Frey et al., 2013). The reasons for these differences in the localisation of tissue cysts of B. besnoiti are poorly understood.

dexamethasone acetate The assessment of the growth rates

The assessment of the growth rates of adult worms as well as SEM evaluations of morphology and dexamethasone acetate of E. canadensis G6 intestinal stages were conducted for the first time in the present work and therefore, comparison with findings from other authors was not possible.
With regard to the growth of rostellar hooks from the metacestode to the adult stage, statistically significant differences were only observed for the LTL and STL. These results agree with and support previous suggestions that such growth is achieved by the addition of new hook material only to certain parts of the hook, whereas the blade appears to be unchanged (Hobbs et al., 1990).
The production of fully developed eggs in E. granulosus sl has generally been regarded to begin between 42 and 48days p.i. in dogs; however, some strains mature earlier (Table 3). Eckert et al. (1989), estimated in “about 40days” the prepatent period for the camel strain based on the observation of worms holding eggs with fully developed embryophores at dexamethasone acetate 41days p.i. The results of the present study confirm that estimation, considering that eggs started to be detected in dog faeces at 41days p.i. and in consequence, the prepatent period was determined as at least, 41days. Moreover, the fact that mature eggs were observed in dog faeces insures that they can effectively reach the environment and infect potential IH on the 41th day after dog infection. Taking into account that the prepatent periods assessed for E. canadensis G6 (camel and goat origin) seem to be identical between them and differ from the 35days period of E. canadensis G7 (pig strain) and of the other species of E. granulosus sl, it is possible that this feature could also be genetically determined by the E. granulosus sl genotype. These results could be taken into account for the design of control strategies based on periodic anthelminthic treatment of dogs, especially in regions where camels are absent but E. canadensis G6 is identified in other IH.

The morphology of adult stages of as well as the prepatent period of the strobilar stage could be genetically determined by the E. granulosus sl genotype. On the other hand, growth and segmentation could be in some way influenced by the IH of origin.
Our findings support the proposal that E. granulosus sl genotypes G6 and G7 should belong to a single species separated from the G8 and G10 genotypes.
The results of this work contribute to increase the knowledge about the biology and genetics of E. granulosus sl complex and are also of practical usefulness for the design of disease control strategies.


Conflict of interest

This study was supported by grant 04/N018 from the Secretaría de Ciencia y Técnica de la Universidad Nacional del Comahue. The authors are grateful to Sebastián Martínez and Bárbara Veuthey for their efficient technical assistance and to Karina Chartier for the collection of parasite samples from rural areas. The authors also wish to thank Patricia Sarmiento for her assistance in the SEM parts of this paper.

Free-roaming domestic cat (Felis catus) populations serve as a valuable resource for studying ectoparasite prevalence. Sometimes called community cats, the term free-roaming can be employed to describe feral, stray, and other non-owned cats which are not considered pets and live exclusively outdoors (Centonze and Levy, 2002). Many of these cats have a close association with homes and human activities, sharing a similar environment as domestic pets. However, since these free-roaming domestic cats have limited or no history of veterinary care or ectoparasiticide use, they provide a unique sampling population to estimate risk of ectoparasite exposure for owned cats under veterinary care.
Ectoparasite studies of free-roaming cats provide value over similarly conducted shelter surveys for multiple reasons. Shelters have varying ectoparasiticide protocols, which may differ within a shelter as well, depending on the individual animal’s personality and response to being handled. Shelter animals are obtained from a variety of sources (i.e., owner surrender, confiscation, stray, or feral) and have an unknown history of acaricide use and veterinary care. Variations in shelter holding time prior to sampling also raises the question of whether parasites were acquired prior to shelter acquisition or from contaminated shelter environments. Similarly, shelter surveys may underestimate the prevalence of tick infestations. Ticks feed for a period of 2–14days (Diamant and Strickland, 1965) and may have fed to repletion and fallen off a host by the time an animal is examined in a shelter survey. Thus, sampling free-roaming domestic cats can provide unique insight into ectoparasite prevalence.

br Acknowledgements The work was supported by

The work was supported by funding from the FP7 GLOWORM project – Grant agreement No 288975CP-TP-KBBE.2011.1.3-04 ( and the Climate Change Research Network (“KLIFF”) by the Ministry for Science and Culture of Lower Saxony (Germany). We thank Dr Brian Boag for useful discussions, and two anonymous reviewers for their useful comments.

Gastrointestinal (GI) nematode infections are considered one of the toughest challenges sheep farmers face worldwide, causing diarrhoea, reduced growth rate, anaemia, and mortality with severe economic losses to individual farmers and the sheep industry as a whole (Hoste and Torres-Acosta, 2011; Mavrot et al., 2015). GI nematode infections used to be controlled with highly effective anthelmintics for several decades, as time and again new products became available while in the meantime anthelmintic resistance (AR) developed to some older products. However, over the last decade, prevalence of AR has risen sharply in the Netherlands with reports on AR to ivermectin, moxidectin and monepantel (Borgsteede et al., 2010; Van den Brom et al., 2013, 2015; Ploeger, unpublished results). A recent review concluded that AR and multi-drug resistance have become widespread in Europe (Rose et al., 2015). These developments have triggered major concerns within the Dutch sheep industry whether current GI nematode control practices are sustainable.
GI nematode control still is based mainly on the use of anthelmintic drugs (Kenyon and Jackson, 2012; Charlier et al., 2014), but it is increasingly recognized that dependency on anthelmintic drugs should be minimized to keep at least some of the drugs effective and, for instance, available for emergency situations. This requires more sustainable control strategies based on grazing management, biological control, host immunity enhancing strategies including vaccination and genetic selection of less susceptible hosts, selective treatment measures and nutritional measures including the use of plants with natural anthelmintic activity (Hoste and Torres-Acosta, 2011). Although increasing knowledge is available on several of these alternative control strategies, acceptance and implementation may not always be an easy process. They have to overcome both farmer’s and veterinary practitioner’s traditional management and perceptions, should be tailor-made aiming at an integrated approach that fits into overall daily management on farm level, and have to be profitable in a relatively short LDN193189 Hydrochloride of time (Van Wyk et al., 2006; Woodgate and Love, 2012). Not every alternative, therefore, may be equally applicable on every sheep farm. Furthermore, specific knowledge gaps on nematode life-cycles and interpretation of, for instance, faecal egg counting results, as well as on utility and applicability of alternative management strategies may hamper implementation by farmers. Finally, implementation of innovative approaches is most likely to occur and sustain when embedded into solid and cooperative social structures (Geels, 2002). In this respect it is of relevance that (1) Dutch sheep farms are partly still under-serviced by veterinary practitioners, (2) the sheep industry is a sector consisting of a variety of sheep farm types with different production goals and not strongly organised as a whole, and (3) that interactive knowledge exchange between parasitology experts, veterinarians and sheep farmers is limited.

Materials and methods


Over the last decades, efforts have been made to change farmer’s and veterinarian’s worm control practices in view of the steadily increasing anthelmintic resistance problems. Main efforts focused on reducing the number of anthelmintic treatments in ewes and lambs, and on pasture management measures such as appropriate rotation schemes to prevent haemonchosis (Eysker et al., 2005). Efforts included posting a web-based treatment advisory tool ( in 2007, making leaflets listing the main worm species and how to control these, and numerous papers in trade journals. Nonetheless, anthelmintic resistance has increased enormously over the last decade, including ivermectin and moxidectin resistance in H. contortus (Borgsteede et al., 2010; Van den Brom et al., 2013, 2015; Ploeger, unpublished results). Moreover, GI nematode infections were recently identified as one of the most challenging and urgent problems by the sheep industry, which has led to funding the present study. Apparently, efforts to educate both farmers and veterinarians are not as effective as one would hope, which conforms to experiences elsewhere (Van Wyk and Reynecke, 2011; Morgan et al., 2012; Woodgate and Love, 2012; McMahon et al., 2013). In order to get handholds for facilitating implementation of alternative strategies, the present survey was conducted to identify sheep farmer’s perceptions concerning worm infections, their anthelmintic treatment practices, their actions to slow down AR development, and their perception of bottlenecks and possible solutions in worm control.

Additionally it seems that administering RBZ alone

Additionally, it seems that administering RBZ alone or in combination with IVM would provide an equivalent therapeutic response, as non significant differences in the overall clinical efficacy (FECR) were found between both treatments. IVM alone failed to control Haemonchus spp. and Cooperia spp., showing efficacies of 0 and 83%, respectively (Table 4). Although Cooperia spp. is known to be the most frequent genus involved in IVM resistance in Argentina (Anziani et al., 2001; Fiel et al., 2001; Suarez and Cristel, 2007), the findings of the present study are in agreement with those of other studies carried out in Argentina, in which both peptide yy were found to be resistant to IVM (Anziani et al., 2004; Fiel et al., 2015). It has been suggested that on farms where resistant nematode populations are present, the use of drug combinations may be an alternative to improve chemical control (Anderson et al., 1988; Geary et al., 2012; Bartram et al., 2012). Although published information in cattle is scarce, some preliminary results indicate that the combination of macrocyclic lactones and LEV was highly effective in minimizing the survival of resistant nematodes (Smith, 2014; Leathwick et al., 2016). Even though the current trial showed no significant improvement in FECR between the RBZ alone and the RBZ+IVM (Table 3), the observed clinical efficacy for the combination was almost as expected, i.e. additive anthelmintic effects between the two drugs (Bartram et al., 2012). An additive effect occurs when the combined effect of two drugs equals the sum of their independent activities measured separately (Entrocasso et al., 2008). This effect has been demonstrated in some trials done in sheep with anthelmintic combinations (Anderson et al., 1991, 1988; Mckenna, 1990b; Entrocasso et al., 2008). In the current study, the marked reduction in the total nematode egg counts 15days after treatment support the high efficacy of RBZ after its administration alone or together with IVM (94% and 98%, respectively). However, it is important to highlight the efficacies against the different genera (Table 4). The efficacy against Cooperia spp. was 83% (IVM), 98% (RBZ) and 98% (IVM+RBZ), while the efficacy against Haemonchus spp. was 0% (IVM), 97% (RBZ) and 100% (IVM+RBZ). Remarkably, the combination was the only treatment that achieved 100% clinical efficacy against IVM-resistant Haemonchus spp. These results are similar to those from a recent field study in cattle, in which efficacies of 78% (IVM), 99% (RBZ) and 99% (IVM+RBZ) against Cooperia spp., and 42% (IVM), 99% (RBZ) and 100% (IVM+RBZ) against Haemonchus spp. were observed (Canton et al., 2015). Once again, the combination was the only treatment that achieved 100% clinical efficacy against IVM-resistant Haemonchus spp. This means that the combined group achieved higher efficacy against resistant parasite populations than did either of the component anthelmintics used alone, with fewer resistant parasites surviving treatment.
The pre-existing level of resistance to one of the anthelmintics in the combination may be a likely explanation for the lack of greater efficacy of the combined treatment. Likewise, Suarez et al. (2014) observed similar nematode control after the use of either a triple combined treatment (LEV+ABZ+IVM) or IVM alone against multiple resistant H. contortus. In sheep, a population of H. contortus not effectively controlled in the field by four anthelmintics (ABM+LEV+ABZ+closantel) administered concurrently in a fixed commercial formulation has also been described (Baker et al., 2012). One of the most important prerequisite criteria to maximize the ability of multiple active formulations by managing existing resistance and slowing its further development is the pre-existing levels of resistance to each of the anthelmintics in the combination. Ideally, the use of nematodicidal combinations may be a valid strategy if the efficacy of each of the anthelmintic molecules approaches 100% (Bartram et al., 2012). This would be the ideal situation to use a nematodicidal combination, but today anthelmintic resistance is unfortunately widespread in Argentina. Therefore, a scenario where the nematode population is susceptible to IVM is not representative of the real situation in most of the commercial cattle farms. As indicated in modeling studies (Dobson et al., 2011; Leathwick, 2012; Leathwick et al., 2012) the key to achieving success with the use of anthelmintic combinations would require there administration before significant resistance (efficacy<70%) to one or more of the active components develops.

Phylogenetic analysis of M and M genes

Phylogenetic analysis of M1 and M3 genes placed TARVs and TERVs into a single genotype while M2 gene divided them into three genotypes. In contrast, phylogenetic analysis of S1 gene (σC protein), which has historically been used to classify ARVs, placed all TARVs in one lineage only (Mor et al., 2014b). Hence, sequencing the M2 gene (μB protein) highlights the importance in understanding the viral EAI045 and relationship within ARVs.
Based on the proposed GCs, the TRVs were divided into three GCs of which GC2 was unique to TRVs only. The maximum number of GCs (n=7) was formed by CRVs of which GC1 and GC3 were shared with TARVs and TERVs, indicating potential reassortments among TRVs and CRVs. An interesting finding was that the maximum number of TARVs (n=9), TERVs (n=5) and all North American CRVstrains (n=5; except strain AVS-B) formed GC1 indicating it to be more prevalent in commercial chicken and turkey populations from North America. Reassortment may be beneficial as it may increase its capacity to replicate in different hosts (Chao, 1997).
The DRVs and GRVs did not share any GC with TARVs, TERVs and CRVs, indicating no reassortments among these viruses. As mentioned earlier, on the basis of sharing of GC1and GC3 of TARVs and TERVs with some CRVs, reassortment between chicken and turkey reoviruses can be expected. The M2-III genotype contained TARV-MN4, TERV-MN5, an enteric CRV strain (AVS-B) isolated in 2006, and an arthritis CRV strain (916S1) isolated in 1992 from Taiwan further indicating possibility of reassortment between chicken and turkey reoviruses. Experimentally, TARVs and TERVs are able to replicate in chickens and may be able to cause enteritis/diarrhea while CRVs are able to replicate in turkeys without causing any disease (Rosenberger et al., 2013a; Sharafeldin et al., 2014).
Co-infections with ARV strains may lead to emergence of new ARV strains, which has been proven in an experimental study by Ni and Kemp (1992). Chicken embryo fibroblasts were co-infected with ARV strain 883 and with 1 of 3 CRV strains (176, S1133, or 81-5), which indicated gene segment selection was virus strain specific. In future studies, co-infections of CRVs, TARVs and TERVs in different combinations will be interesting to understand reassortment events among these viruses.
TERVs have been isolated from apparently healthy and enteritis-affected turkeys for years (Pantin-Jackwood et al., 2008; Jindal et al., 2010, 2014) but, until recently, there have been no reports on reovir### lameness and arthritis in turkeys after it was first reported in the 1980s (Levisohn et al., 1980; Al Afaleq and Jones, 1989). After a hiatus of >20 years, the problem of turkey arthritis re-emerged in the upper Midwest area initially and then was reported from several other US states in both commercial and breeder turkey flocks. Currently, the TARV-Crestview strain is used as an inactivated, autogenous vaccine in the field (Dave Mills, personal communication). When this vaccine strain was compared to TARVs and TERVs, we found two (M436W, D437N) and one (L260F) unique aa residues in μA and μB proteins, respectively, differentiating the vaccine from wild strains. It is not known whether TARV-Crestview strain accords full protection against TERVs. Further studies are needed in this area.
Recently, new variants of CRVs associated with viral arthritis in broiler chickens have been reported from Europe and North America (Rosenberger et al., 2013a; Sellers et al., 2013; Troxler et al., 2013). Based on virus neutralization test, these variants of CRVs and TARVs were found to be antigenically different and did not cross neutralize (Rosenberger et al., 2013b). In addition, commercially available CRV vaccines were found to be ineffective against these new CRV variants (Sellers et al., 2013; Troxler et al., 2013). The occurrence of these two events simultaneously in two different host species (chickens and turkeys) raises the question on the source of these arthritis-associated reoviruses. Unfortunately, sequences from these new CRV variants were not available for comparison in the GenBank. There could be three possibilities for the sudden appearance of these viruses in turkeys: firstly, sequencing data could indicate that CRV strains are closely related to TRV strains and share one genomic constellation (GC1), which probably suggests possible common source of TARV and new CRV variants, but turkey and chicken hatcheries do not share a common connection. However, possible virus transmission between these two types of units may occur via aerosols, wild birds, or by mechanical means (e.g., through fomites, personnel, or farm equipment).

In compliance with the fast spreading

In compliance with the fast spreading of the virus within carp (Gilad et al., 2004), remarkable haematological changes occurred in CyHV-3 infected carp. Already by day 2 post infection, infected carp experienced anaemia and a pronounced leucocytosis, characterised by increased proportions of granulocytes and monocytes. These cellular indicators for an acute inflammation confirm previous findings of an escalation of C-reactive protein (CRP) and alternative complement activity levels immediately after exposure of carp to the virus (by 6h post infection, Ponnier et al., 2014) indicating a strong and rapid acute phase response in these fish. The immediate response of the acute phase proteins, as seen by Ponnier et al. (2014) and the haematological changes (anaemia and leucocytosis) observed in the present study are remarkable, since at this point no or minimal pathological changes were recorded. However, data on virus load and replication from previous publications (Adamek et al., 2013, 2014; Gilad et al., 2004; Miwa et al., 2014) indicate that the virus spreads throughout the whole body and reached multiple organs within a short period of time after exposure to the pathogen. This leads to a significant up-regulation in the BLU9931 of several immune related genes, including genes involved in the regulation of acute phase protein expression (Rakus et al., 2012). An increased vascular permeability, which is associated with acute inflammation, may intensity solute movement across the branchial epithelium and by this reinforce the osmotic dysfunction in CyHV-3 infected carp.


Papillomaviruses (PVs) are small circular double-stranded DNA viruses. As PVs can influence cell growth and differentiation, some are important causes of neoplasia. Papillomaviruses are classified into genera based on the sequence of the highly conserved ORF L1 (Bernard et al., 2010). While the overwhelming majority of PVs only infect epithelium and are highly host specific (Bernard et al., 2010; Joh et al., 2011), the bovine papillomaviruses (BPVs) of the Deltapapillomavirus genus have the ability to infect both epithelial and mesenchymal cells and to infect multiple species (Munday, 2014).
Compared to most species PV-induced disease is currently thought to be rare in cats. Diseases that are recognised to be caused by PVs in domestic cats include oral papillomas due to Felis catus (Fca) PV-1 (Munday et al., 2015), feline cutaneous viral plaques and Bowenoid in situ carcinomas due to infection with FcaPV-2 and FcaPV-3 (Lange et al., 2009; Munday et al., 2013, 2008), and feline sarcoids. Feline sarcoids are BLU9931 rare mesenchymal neoplasms that most commonly develop around the face and digits of younger domestic cats (Schulman et al., 2001), but have also been reported in mountain and African lions (Orbell et al., 2011; Schulman et al., 2003). While metastasis has not been reported, these neoplasms are progressive and can result in euthanasia due to recurrence following surgical excision. The same PV DNA sequence, designated the feline sarcoid-associated PV sequence (FeSarPV), has been consistently detected in multiple studies of sarcoids from domestic cats and exotic felids (Munday et al., 2010; Orbell et al., 2011; Teifke et al., 2003). As the FeSarPV sequence only comprises a short segment of the PV L1 gene, classification of this PV has not been possible. However, as FeSarPV cannot be amplified from any non-sarcoid feline sample (Munday et al., 2010) but can be amplified from samples of normal skin and fibropapillomas of cattle (da Silva et al., 2012; Munday and Knight, 2010) it appears likely that, similar to equine sarcoids, feline sarcoids are due to cross-species infection by a BPV type (Orbell et al., 2011; Schulman et al., 2001). The aim of the present study was to amplify the entire genome of the PV that contains the FeSarPV sequence. Classification and phylogenetic analysis of this PV could then be used to support a bovine definitive host.

SCR7 The increased replication of PCV

The increased replication of PCV2 observed in this study due to environmental stress being placed on the pigs can be attributed to a few factors. Firstly, pigs kept in high stocking density are more likely to come into contact with any infected pigs already in the pen and therefore are more regularly exposed to virus. Secondly, increased stress on the pig as an individual can lead to increase in serum levels of the stress hormone cortisol. This increase in serum cortisol levels has been demonstrated in pigs in response to food and water deprivation (Parrott et al., 1989), heat stress (Becker et al., 1985), social stress (Parrott and Misson, 1989a) and shipping of animals (McGlone et al., 1993; Nyberg et al., 1988; Parrott and Misson, 1989b). Higher levels of serum cortisol associated with a higher level of stress have been demonstrated to be associated with a reduced natural killer (NK) cell cytotoxicity as well as a reduced number of circulating lymphocytes in the blood (McGlone et al., 1993). As NK SCR7 are an essential component of the innate immune system and are heavily involved in the early response to viral infection, the increased PCV2 replication observed in stressed pigs in this study may be explained by a similar inhibition in NK cell function. Further study into the cortisol levels and NK cell function in the system described in this study would indeed be interesting and could potentially further develop the PCV2 infection model.
Results from the present study showed that on average, PCV2 copies in the mesenteric lymph node, lung and bone marrow are higher in pigs from the V SD T group compared with those subjected to only one environmental stress or C pigs. In the inguinal lymph node however, it was observed that the average PCV2 copy number was highest in the V SD group. This high average was a result of one animal having a very high PCV2 copy number in this tissue. Although the daily behaviour of the pigs was not monitored, such high PCV2 copy number in the inguinal lymph node of a single pig may be a result of that pig being submissive. McGlone et al. found increased levels of serum cortisol and an associated decrease in NK activity in pen mates that were determined submissive compared with those that were determined dominant (McGlone et al., 1993).

This work was funded by a grant (BB/FO18394/1) from the BBSRC ‘Combating Endemic Diseases of Farmed Animals for Sustainability’ (CEDFAS) initiative, with contributions from BPEX, Biobest Laboratories and Zoetis Animal Health. We thank the staff of the Biological Service Unit of the RVC, especially A. Wallis and C. Davies for their technical help. This manuscript represents publication number PPB_00864 of the RVC.

Iridoviridae, a large icosahedral enveloped viruses present in the cytoplasm were divided in### five genus: Iridovirus, Chloriridovirus, Lymphocystivirus, Ranavirus, Megalocytivirus (Jancovich et al., 2012). Iridoviruses were well known as causative agents of serious systemic diseases among feral, cultured, and ornamental fish in the last decade worldwide (Wang et al., 2007). Among family Iridoviridae, members of genus Lymphocystivirus, Ranaviruses and Megalocytiviruses affected finfish. Both ranaviruses and megalocytiviruses cause severe systemic disease, occur globally and affect a diversity of hosts. Ranaviruses are also significant pathogens of amphibians. In contrast, lymphocystiviruses, although widespread in fish, rarely cause economic loss (Whittington et al., 2010). The genus Megalocytivirus included red sea bream iridovirus (RSIV), infectious spleen and kidney necrosis virus (ISKNV), turbot reddish body iridovirus (TRBIV), dwarf gourami iridovirus (DGIV), Taiwan grouper iridovirus (TGIV), Sea bass iridovirus (SBIV) and rock bream iridovirus (RBIV), which caused significant mortality in multiple species of marine and freshwater fish (Inoue et al., 1992; Kurita and Nakajima, 2012; Shuang et al., 2013). Histopathological features of genus Megalocytiviruses were the formation of distinctive hypertrophied cells sometimes in large numbers throughout various organs, especially spleen (Whittington et al., 2010). The frog virus 3 (FV3), epizootic haematopoietic necrosis virus (EHNV), European catfish virus (ECV), largemouth bass virus (LMBV), Singapore grouper iridovirus (SGIV) and grouper iridovirus (GIV) were classified into genus Ranaviruses, which caused severe necrosis to internal organs of many fishes, especially in spleen and renal haematopoietic tissue (Ahne et al., 1989; Chao et al., 2002; Chinchar, 2002; Langdon and Humphrey, 1987; Langdon et al., 1988, 1986; Murali et al., 2002; Pozet et al., 1992; Plumb et al., 1996, 1999; Qin et al., 2003). More and more evidences showed that ranavirus have become a significant cause of disease in ectothermic animals, and that form a virological, commercial and ecological point of view deserve additional study (Chinchar, 2002).

Conversely we observed that H parasuis significantly decreased the

Conversely, we observed that H. parasuis significantly decreased the level of ROS detected in supernatants of infected PAMs (Fig. 3). The reduction in ROS was confirmed using also other strains of H. parasuis (serotype 4, serotype 6) (data not shown). Many bacterial species colonising the respiratory tract have evolved detection and response systems to prevent damage by ROS. For example, Haemophilus influenzae possesses an anti-ROS defence mechanism mediated by catalases and superoxide dismutase production (Whitby et al., 2012 and Eason and Fan, 2014). Haemophilus parasuis is known for its resistance to CAL101 (Costa-Hurtado et al., 2012) and, based on our results, we speculate that an analogous anti-ROS defence mechanism exists in H. parasuis.

The present study provides a description of the immune response of PAMs to a dual respiratory infection of swine. Co-infection of PAMs with PRRSV and H. parasuis elicited a pro-inflammatory immune response represented by significant IL-1β production. Infection of PAMs with H. parasuis was associated with decreased production of ROS, indicating the presence of an H. parasuis defence mechanism against respiratory burst. Data obtained in this study show that multifactorial respiratory disease in natural conditions associated with PRRSV and H. parasuis infections could be the consequence of pro-inflammatory mediated immunopathology.

Competing interests


Schmallenberg virus (SBV) is an arbovirus of the Orthobunyavirus genus that is transmitted by biting midges (Culicoides spp.). Since its identification at the end of 2011 (Hoffmann et al., 2012), SBV has spread rapidly throughout mainland Europe. SBV infection of adult ruminants appears to be sub-clinical or mild; causing watery diarr###, fever and reduced milk production (Muskens et al., 2012). However, infection of animals during pregnancy causes arthrogryposis–hydranencephaly syndrome (AHS), which results in congenital malformations, abortions and stillbirths (Tarlinton et al., 2012).
Although the original identification of SBV infection was made following observation of acute signs in adult dairy cattle from late summer 2011 (Hoffmann et al., 2012), the majority of SBV infections are reported due to the appearance of AHS in calves and lambs. The first cases of AHS were reported in the Netherlands in November and December 2011 and in Belgium in 2012 (Garigliany et al., 2012; van den Brom et al., 2012). By comparison with the related Akabane virus (Kirkland et al., 1988), it is suspected that SBV causes AHS only if infection occurs in the mid-stages of pregnancy (Tarlinton et al., 2012). Therefore, it is assumed that when AHS is observed, SBV must have been circulating several months previously. This is supported by the initial detection of SBV in France in January of 2012 on the basis of malformed lambs (Dominguez et al., 2012) with subsequent retrospective analysis identifying seropositive animals sampled in October 2011 (Zanella et al., 2013). Studies of Belgian ruminants found that almost all animals were seropositive for SBV at the end of 2011 (Meroc et al., 2014, 2013a). Although the duration of acquired immunity for SBV remains unknown, it was speculated that herd immunity would prevent a second epidemic in 2012. In a follow-up study, anti-SBV antibody titres remained high in animals one year later and very few clinical cases were reported in 2012 (Meroc et al., 2013b). SBV infection in the United Kingdom (UK) was first identified in malformed lambs from farms in south-eastern coastal regions (Kent, East Sussex, Norfolk and Suffolk) in January 2012 (APHA, 2012; Roberts, 2012).

Materials and methods
Virus neutralisation tests (VNT) were carried out as described in Loeffen et al. (2012) using virus strain BH80/11-4 (species Schmallenberg virus, genus Orthobunyavirus, family Bunyaviridae) (kindly provided by M. Beer, Friedrich–Loeffler Institut) with the minor modification that cells were fixed by the addition of 100% ethanol and stained using 0.1% v/v methylene blue in water. Positive and negative controls (samples previously tested with the SBV IDscreen indirect ELISA [IDvet, France] by [BioBest Laboratories, UK]) were tested in parallel with every batch of VNTs.