A cross-sectional study of bluetongue virus and Mycoplasma bovis infections in dairy cattle: II. The association between a positive antibody response and reproduction performance

In a cross-sectional study to determine the possible relationship between a positive antibody test to bluetongue virus (BTV) or Mycoplasma bovis infections and reproductive performance of dairy cows, data were collected on 572 California dairy cows during December 1986 for analysis. Serum samples were tested using an enzyme-linked immunosorbent assay (ELISA). Data on reproduction variables were extracted from the individual cow sheets of the California Dairy Herd Improvement Association records and interfaced with the serological results for analysis.Similar data analyses for both BTV and M. bovis were performed to identify and quantitatively assess the association of the reproduction variables and each agent. These associations were evaluated unconditionally using the x ² for categorical variables and Student's t-test for continuous variables. Logistic regression analysis was used to determine if reproduction variables with significant unconditional associations remained significant when adjusted for the effects of possible confounding factors.Both the BTV and M. bovis ELISA antibody titres indicated exposure to the agents. The results of the multiple logistic regression analyses indicated that cows seropositive for BTV were significantly older at first calving (p<0.03). For M. bovis, seropositive cows were more likely to have longer intervals from calving to last service and longer intervals from calving to pregnancy diagnosis than seronegative cows (p<0.05). The other reproduction variables examined were not significantly associated with ELISA seropositivity.     

Association between risk of seroconversion of sentinel cattle to bluetongue viruses and Culicoides species (Diptera: Ceratopogonidae) in Queensland, Australia

The association between risk of seroconversion of sentinel cattle to bluetongue viruses and the number of Culicoides brevitarsis Kieffer and C. wadai Kitaoka caught by light traps was investigated using survival analysis. Eight sentinel herds that seroconverted to bluetongue viruses between 1990 and 1994, and for which insect-trapping data were available, were selected for inclusion in the study. These herds were located at six sites along the eastern coast of Queensland, Australia, from approximately latitude 10 °South to 25 °South. C. brevitarsis was detected at all locations where sentinel herds were maintained, whereas C. wadai was detected at only two locations in northern Queensland where four sentinel herds were maintained during the study period. The mean number of C. brevitarsis and C. wadai caught per month was 230 and 21, respectively. A significant (P = 0.05) positive association was found between the risk of seroconversion of sentinel cattle to bluetongue viruses and the number of C. wadai caught in the same month.     

A cross-sectional study of bluetongue virus and Mycoplasma bovis infections in dairy cattle: I. the association between a positive antibody response and production efficiency

In a cross-sectional study of bluetongue virus (BTV) and Mycoplasma bovis (M. bovis) infections, a sample of 572 California dairy cows was tested for the presence of antibodies to answer the question: Is it possible to identify and to assess quantitatively the associations between positive antibody test and production? Serum samples collected from these cows during December 1986 were tested for the presence of antibodies to BTV and M. bovis using the enzyme-linked immunosorbent assay (ELISA). Data on milk production were extracted from individual cow sheets of the California Dairy Herd Improvement Association (DHIA) record-keeping system and interfaced with percentage ELISA results for analysis. Univariate and multivariate statistical analyses, using the X ² test for categorical variables or Student's t-test for continuous variables and multiple logistic regression respectively, were carried out to evaluate for possible associations between positive antibody tests to each agent and each production variable of interest. Complete data on all variables studied were obtained for 289 (50.5%) cows for M. bovis and 423 (74%) cows for BTV. For cows with complete data on all variables, estimates of the point prevalence of antibodies to BTV and M. bovis were 70.5% and 66.1%, respectively. Results of this study indicated that Guernsey cows were more likely to have a positive BTV test than Holstein cows and that cows in higher lactations were more likely to test positive to BTV ELISA than those in lower lactations (p<0.05). Because all cows except those on one farm were Holstein, our confidence in the effect of breed is limited. The association between lactation number and BTV seropositive test may be an age factor identified earlier in the study. For M. bovis, the results of the analysis indicated that seropositive cows were more likely to produce less milk, on a mature equivalent basis (OR_adj=0.96, p=0.034), and that they had less extended 305 day milk production potential (OR_adj=0.90, p<0.0001) than seronegative cows.     

A seroepidemiological study on bluetongue virus in dairy cattle in the central valley of California

A seroepidemiological study on bluetongue virus (BTV) infection in California dairy cattle was conducted to estimate the prevalence and distribution by age and season of BTV group-reactive antibodies and to look for possible associations between the presence of antibodies and cattle age or breed and farm. Between December 1985 and March 1987, a sample of cattle was tested at approximately two-month intervals for BTV group-reactive antibodies using an enzyme-linked immunosorbent assay (ELISA). Data taken during the month of December 1986 were used to evaluate possible associations between a positive antibody test and certain intrinsic (age, breed) and extrinsic (farm) factors.Univariate and multivariate statistical analyses using the -square test for associations and multiple logistic regression, respectively, were carried out for possible associations between positive antibody tests to BTV and each factor of interest. The strengths of the associations were determined using estimates of the odds ratio.Of the 3774 serum samples tested, 238 (6.3%) were from calves, 1045 (27.6%) were from heifers and 2492 (66.0%) were from cows. Seroprevalence varied from nil in calves on two occasions to over 90% on several occasions in cows. Cows consistently had higher prevalence rates than heifers or calves across all test dates (p<0.05). The seroprevalence of BTV group-reactive antibodies also showed a seasonal fluctuation, with the highest rates occurring during the warmer months of the year. These highest prevalence rates coincided with heavy activity of the known vector of BTV, Culicoides spp. Breed and farm effects were not statistically significant (p>0.05). With the exception of one farm, all cattle were of the Holstein breed, which reduced confidence in assessing any breed effect in this study. Relative estimates of the sensitivity and specificity of BTV ELISA were 87% and 100% respectively, compared to the standard agar gel immunodiffusion (AGID) test.The observations support previous findings of seasonal distribution of BTV antibodies and suggest an age relationship, whereby older cattle are more likely to be positive to BTV group-reactive antibodies than younger cattle.     

Spatial distribution and habitat selection of culicoides imicola: The potential vector of bluetongue virus in Tunisia

The increasing threat of vector-borne diseases (VBDs) represents a great challenge to those who manage public and animal health. Determining the spatial distribution of arthropod vector species is an essential step in studying the risk of transmission of a vector-borne pathogen (VBP) and in estimating risk levels of VBD. Risk maps allow better targeting surveillance and help in designing control measures. We aimed to study the geographical distribution of Culicoides imicola, the main competent vector of Bluetongue virus (BTV) in sheep in Tunisia. Fifty-three records covering the whole distribution range of C.imicola in Tunisia were obtained during a 2-year field entomological survey (August 2017 – January 2018 and August 2018 – January 2019). The ecological niche of C. imicola is described using ecological-niche factor analysis (ENFA) and Mahalanobis distances factor analysis (MADIFA). An environmental suitability map (ESM) was developed by MaxEnt software to map the optimal habitat under the current climate background. The MaxEnt model was highly accurate with a statistically significant area under curve (AUC) value of 0.941. The location of the potential distribution of C. imicola is predicted in specified regions of Tunisia. Our findings can be applied in various ways such as surveillance and control program of BTV in Tunisia.     

Development and evaluation of a real-time quantitative PCR assay for Culicoides imicola, one of the main vectors of bluetongue (BT) and African horse sickness (AHS) in Africa and Europe

The current microscopy method for identifying the Culicoides imicola Kieffer, 1913 species can be time and labour intensive. There is a need for the development of a rapid and quantitative tool to quantify the biting midges C. imicola ss in light trap catches. A reproducible and sensitive real-time polymerase chain reaction method that targets the internal transcribed spacer (ITS-1) of ribosomal DNA of C. imicola ss species was developed. This real-time PCR assay was first performed on 10-fold serial dilutions of purified plasmid DNA containing specific C. imicola ss ITS-1. It was then possible to construct standard curves with a high correlation coefficient (r2=0.99) in the range of 10(-2)-10(-8) ng of purified DNA. The performances of this PCR were evaluated in comparison with morphological determination on Culicoides trapped along the Mediterranean coastal mainland France. ROC statistical analysis was carried out using morphology as gold standard and the area under the ROC curve had a satisfactory value of 0.9752. The results indicated that this real-time PCR assay holds promise for monitoring C. imicola ss population in both surveillance and research programmes because of its good specificity (92%) and sensitivity (95%).     

The distribution ofParascaris equorum eggs in the soil profile of bare paddocks in some Norwegian studs

Fifteen bare paddocks, which consisted of soil only and were located on 12 different studs, were examined for their content ofParascaris equorum eggs in the upper 15 cm of the soil profile. The paddocks were classified into three different groups according to the type of soil: clayey soil (A), morainic soil (B) and gravel or gravel-like sand (C). Soil profiles were collected down to a depth of 15 cm and were divided into three layers: 0–5 cm (D1), 5–10 cm (D2) and 10–15 cm (D3). The eggs in each layer were counted and identified as infective or noninfective eggs. The paddocks in group C, which had good drainage conditions, had significantly lower numbers of eggs in the whole profile and in D1 and D2 than the paddocks in groups A or B. Furthermore, there was a significantly higher proportion of the total egg count present in D3 in the group C paddocks. This may have been due to a higher degree of passive transportation of eggs down the profile in the gravel or gravel-like sand. Even though there was a significantly higher proportion of infective eggs in the soil from the group C paddocks, the lower total numbers of eggs resulted in a lower total number of infective eggs in those paddocks. The study showed that the soil type was an important factor in determining the content ofP. equorum eggs in the upper layer of the soil profile in bare paddocks and consequently for the potential infestation of horses withP. equorum.     

Seasonal dynamics of biting midges (Diptera: Ceratopogonidae: Culicoides), the potential vectors of bluetongue virus, in Sweden

The outbreak of bluetongue (BT) in northern Europe 2006 initiated the monitoring of vectors, biting midges of the genus Culicoides in Sweden. In order to determine the diversity, distribution and seasonal dynamics of Culicoides, weekly collections were made during 2008 and during March-December 2009 using the Ondestepoort Veterinary Institute black light trap. Twenty sampling sites were selected in 12 provinces. In total of 30,704 Culicoides were collected in 2008 and 32,252 in 2009. The most abundant species were the potential vectors of BTV Culicoides obsoletus/C. scoticus that comprised of 77% of the total catches. Other biting midges collected were Culicoides impunctatus (9%), Culicoides grisescens (3%), Culicoides punctatus (2%), Culicoides chiopterus (2%) and Culicoides pulicaris (2%). Culicoides obsoletus/C. scoticus were most abundant during May-June and August-September. The majority of the species were active from March to November in 2008 and April to October in 2009. Species considered as potential vectors of bluetongue virus (BTV) occurred as far north as latitude 65°N (Kalix).     

The distribution pattern of Sarcocystis species,their transmission and pathogenesis in sheep in Fars Province of Iran

Of 1362 sheep examined during two years in Fars Province of Iran, 786 (57.7%) were positive for Sarcocystis spp. The prevalence was significantly higher (p<0.05) in animals owned by nomadic Assyrians (67.95%) than in those owned by local people (41.86%). More of the animals above 2 years age were infected (69.98%) than young ones (30.02%). Females had a higher prevalence of infection (61.07%) than males (38.93%) but most of the males were younger. There was no variation in the infection rate during spring, summer or autumn, but it was low in winter. The species observed were Sarcocystis gigantea, predominantly in oesophagus, S. medusiformis, mainly in diaphragm, S. tenella in the oesophagus, diaphragm, tongue and heart, and S. arieticanis in the oesophagus, tongue and occasionally in the diaphragm. In transmission studies, the prepatent period for S. gigantea and S. medusiformis and for the two microscopic species was 11–13, 10 and 8–12 days, respectively. The infection could not be transmitted to hamsters and guinea-pigs. The macroscopic species were almost non-pathogenic but were responsible for economic losses because of rejection of carcases or parts thereof at slaughter. The microscopic species caused tissue damage to the affected organs, resulting in haemorrhages, mononuclear infiltration and necrotic changes.Abbreviations DPI days post infection     

The 2006 outbreak of bluetongue in northern Europe—The entomological perspective

After bluetongue (BT) appeared in northern Europe in August 2006 entomological studies were implemented in all five affected Member States (MSs) to establish which species of Culicoides had acted as vectors. The findings can be summarised as follows: (i) C. imicola the principal southern European/African vector of BTV has not penetrated into northern Europe, (ii) three pools of C. obsoletus/C. scoticus and one of C. dewulfi assayed RT-PCR-positive to BTV-8, (iii) in support of these results it was found that both potential vectors had also high parity rates (approximately 40%) indicating increased longevity favouring BTV virogenesis and transmission, (iv) furthermore, C. obsoletus/C. scoticus and C. dewulfi occurred also widely and abundantly on sheep and cattle holdings across the entire affected region, (v) and during the latter part of the season showed strong endophily readily entering livestock buildings in significant numbers to bite the animals inside (endophagy), (vi) which demonstrates that housing at best offers only limited protection to livestock from Culicoides attacks, (vii) in contrast the potential vector C. pulicaris sensu stricto was restricted geographically, was captured rarely, had a low parity rate (10%) and was exophilic indicating it played no role in the outbreak of BT, (viii) the incrimination of C. dewulfi as a novel vector is significant because it breeds in cattle and horse dung this close association raising its vectorial potential, but (ix) problems with its taxonomy (and that of the Obsoletus and Pulicaris species complexes) illustrates the need for morphological and molecular techniques to become more fully integrated to ensure progress in the accurate identification of vector Culicoides, (x) midge densities (as adjudged by light traps) were generally low indicating northern European Culicoides to have a high vector potential and/or that significant numbers of midges are going undetected because they are biting (and transmitting BTV) during the day when light traps are not effective, and (xi) the sporadic capture of Culicoides in the winter of 2007 invites re-examination of the current definition of a vector-free period. The re-emergence of BT over a wide front in 2007 raises anew questions as to precisely how the virus overwinters and asks also that we scrutinise our monitoring systems in terms of their sensitivity and early warning capability.     

The distribution of invA,pagC and spvC genes amongSalmonella isolates from animals

New molecular diagnostic techniques often rely on hybridization or amplification of specific DNA regions to detect pathogenic bacteria. The choice of genes to be used as probes or as the targets of amplification techniques is critical to the success of these procedures. The genes so used might best be those associated with virulent isolates and having a wide distribution among such isolates. In this study three genes,invA, pagC andspvC, thought to be associated with the virulence of salmonellae, were labelled and used to probe the total DNA from 103Salmonella isolates from animals in an attempt to determine whether these genes might be useful in diagnostic procedures.pagC was detected in 99% of theSalmonella tested, andinvA was detected in 94.2% of the isolates. BothpagC andinvA were detected with a significantly higher frequency thanspvC in isolates from chickens and swine, but no significant difference in detection of these three genes occurred when bovine isolates were examined. Failure to detect any of these genes occurred in only one isolate. Isolates from apparently healthy or from clinically ill chickens and swine could not be distinguished by detecting these three genes. The genes were not detected in the non-Salmonella strains tested. These results suggest that, of these three genes,pagC may be the best choice for use as a probe or polymerase chain reaction target in future detection protocols.Abbreviations df degrees of freedom - H apparently healthy animal - I animal diagnosed clinically as having salmonellosis - LB Luria-Bertani - NDSU North Dakota State University in Fargo, North Dakota - NS not significantly different - NVSL National Veterinary Services Laboratory in Ames, Iowa - PCR polymerase chain reaction - SDS sodium dodecylsulphate - UGA University of Georgia in Athens, Georgia     

Gastrointestinal helminths and lungworms of French dairy goats: Prevalence and geographical distribution in Poitou-Charentes

A study was conducted on 81 dairy-goat farms in western France. Faecal samples were collected once and 31 culled goats were necropsied in order to assess their gastrointestinal and respiratory helminth species, their prevalences and the intensities of infection as well as their geographical distribution in six subregions of the surveyed area. Fifteen species of helminth were recovered. The most frequent wereMuellerius capillaris, Trichostrongylus colubriformis andTeladorsagia circumcincta with a prevalence more than 90%. The two latter species represented respectively 50.9% and 30.2% of the total worm burden of the digestive tract. Other prevalent species wereHaemonchus contortus, Strongyloides papillosus, Trichuris spp.,Moniezia spp. andDicrocoelium lanceolatum. The occurrence ofDicrocoelium lanceolatum was related to the presence of calcareous or alkaline soils and restricted to the south of the surveyed area. No area at particular risk was defined for the other helminth infections, probably because of the relative homogeneity in climatic and topographic conditions of the six agricultural regions.     

Prevalence of agglutinating antibodies to Toxoplasma gondii in small ruminants of the Madrid region,Spain, and identification of factors influencing seropositivity by multivariate analysis

A seroepidemiological survey of Toxoplasma gondii infection in sheep and goats was conducted in the Madrid region of Spain. Sera were collected from 60 herds, for which farming management information and other relevant data for their characterization were also obtained through a questionnaire. The seroprevalence was 11.8% (64 out of 541), using the modified (2-mercaptoethanol) direct agglutination technique with a 1:64 cut-off titre. The relationship between seropositivity and the variables in the questionnaire was assessed by multivariate analysis. Four variables were found to be significantly associated with seroprevalence. Two of them, the presence of cats and a previous history of abortion outbreaks in the farm, were factors known to be linked with toxoplasmosis, indicating the validity of the serological data. Seropositivity was also related to a lack of replacements in the preceding year. Proximity to other farms appeared to be a protective factor negatively associated with seropositivity, probably because it was an indicator of proximity to an urban area and the availability of local sanitary facilities.Abbreviations AIDS acquired immune deficiency syndrome - logistic regression coefficient - CI confidence interval - MDAT modified direct agglutination test - OR odds ratio - p level of significance - SE standard error     

Simulation analysis of the effect of herd immunity and age structure on infection of a cattle herd with bluetongue viruses in Queensland, Australia

A state-transition model based on Leslie matrix formulation was used to investigate the effects of herd immunity and age structure on the infection of a simulated cattle herd with bluetongue viruses under Australian climatic conditions. Increasing duration of immunity decreased the prevalence of infection. A duration of immunity of 33 months was consistent with prevalence estimates made from previous serological studies of bluetongue virus. Herd prevalence displayed slowly dampening cyclical variation over time (most pronounced when a short duration of immunity was simulated). Increasing calving and mortality risk rates in the simulated herd increased prevalence, whereas increasing age at first calving decreased prevalence. Manipulation of calving rates had the greatest effect on the predicted prevalence of infection in the herd. Simulation of a number of herd-management scenarios suggested that management systems in which cattle are bred early and where high calving rates are achieved are likely to contribute to high levels of infection with bluetongue viruses. Results confirm the importance of management factors in influencing the prevalence of infectious diseases in animal populations.     

Seasonal Variation of Boophilus microplus (Canestrini, 1887) (Acari: Ixodidae) in Cattle in Minas Gerais State,Brazil

The seasonal variation of the tick Boophilus microplus in the `Regiaõ Metalúrgica' region of Minas Gerais state was studied using 26 Bos taurus (Holstein)×Bos indicus (Gir) calves, of which 16 animals were 3/4 Holstein×1/4 Gir and 10 were 3/4 Gir×1/4 Holstein. Monthly tick counts were made between March 1996 and March 1998 and, although B. microplus infestations occurred throughout the year, they were greater during the rainy season. Rainfall was the climatic factor that most affected the seasonal variation. The animals with a preponderance of Holstein blood presented infestations that were significantly (p<0.05) greater than those of the other cross. Five of the animals (19.2%) maintained 45.5% of the parasite load of the entire group.     

Indoor activity of Culicoides associated with livestock in the bluetongue virus (BTV) affected region of northern France during autumn 2006

In August 2006, bluetongue virus (BTV) was detected in the Netherlands, Belgium, western Germany, Luxembourg and northern France for the first time. Consequently, a longitudinal entomological study was conducted in the affected region of northern France (Ardennes) throughout the autumn of 2006. Data on the spatio-temporal distribution of Culicoides (Diptera: Ceratopogonidae) associated with livestock were collected and an attempt was made to identify the vector(s) involved in BTV transmission by means of virus detection in wild-caught biting midges. Weekly sampling using standardized Onderstepoort-type blacklight traps were performed simultaneously both outdoors and indoors in one BTV-free and three BTV-affected farms between September and December 2006.Culicoides were sorted according to farm, location (outdoors vs. indoors), time point (in weeks), species and physiological stage. BTV detection was conducted by RT-PCR on monospecific pools of non-bloodfed parous female Culicoides.The principal results showed: (i) the absence of the Mediterranean vector, C. imicola, (ii) the relatively low abundance of C. dewulfi and C. pulicaris, (iii) the widespread occurrence and abundance of C. obsoletus/C. scoticus with longevity and behaviour compatible with BTV transmission, and (iv) all Culicoides pools tested for BTV were negative.In France, the very low levels of BTV-8 circulation were probably due to the limited introduction of the virus from affected neighbouring countries, and not due to the absence of local vector populations. A key finding has been the substantiation, for the first time, that Culicoides, and particularly the potential vectors C. obsoletus/C. scoticus and C. dewulfi, can be active at night inside livestock buildings and not only outside, as originally believed.The endophagic tendencies of members of the Obsoletus group are discussed in light of the prolonged period of BTV transmission during the autumn of 2006 and the risk of BTV overwintering and resurgence in the spring of 2007. Overall, there is an urgent need to improve our knowledge on the ecology of local Culicoides species before any clear, effective and reliable recommendations can be provided to the veterinary authorities in terms of prevention and control.     

The distribution of Fasciola hepatica in Queensland, Australia, and the potential impact of introduced snail intermediate hosts

A survey was conducted to establish the distribution of the liver fluke, Fasciola hepatica, in the state of Queensland, Australia, and to evaluate the impact of the introduced snail intermediate hosts, Pseudosuccinia columella and Austropeplea viridis. Serum samples from a total of 5103 homebred cattle in 142 beef herds distributed throughout the state and 523 pooled milk samples from dairy herds from the state's major dairying regions were tested for antibodies to F. hepatica by ELISA. Snails were collected on infected properties around the limits of the F. hepatica distribution. F. hepatica infection was detected in 44 dairy herds and two beef herds. The distribution of infected herds indicates that F. hepatica is established only in southeast Queensland. The distribution there was patchy but the parasite was more widespread than suggested by an earlier survey. The predominant intermediate host species found along the northern limit of the distribution was P. columella. We conclude that the introduction of P. columella and A. viridis has not yet had a major impact on the distribution of F. hepatica in Queensland. However, the presence of P. columella, which is much more adaptable to tropical habitats than the native intermediate host, Austropeplea tomentosa, at the northern limit of the F. hepatica distribution suggests that there is potential for the parasite to expand its range.     

Ecological correlates of bluetongue virus in Spain: predicted spatial occurrence and its relationship with the observed abundance of the potential Culicoides spp. vector

Using data from bluetongue (BT) outbreaks caused by viral serotype 4 (BTV-4) in Spain during 2004–2005, a predictive model for BTV-4 occurrence in peninsular Spain was developed. An autologistic regression model was employed to estimate the relationships between BTV-4 presence and bioclimatic-related and host-availability-related variables. In addition, the observed abundances of the main potential Culicoides vectors during 2004–2005, namely Culicoides imicola, Culicoides obsoletus group, and species of the Culicoides pulicaris group, were compared between BTV-4 presence/absence areas predicted by the model.BTV-4 occurrence was mainly explained by bioclimatic variables, although a consideration of host-availability variables led to improved fit of the model. The area of BTV-4 presence predicted by the model largely resembled the core distribution area of C. imicola, and this species was the most abundant Culicoides spp. in predicted BTV-4 presence areas. The results suggest that the spatial expansion of BTV-4 took place only as far as those areas in which C. imicola populations efficiently transmitted the virus.     

Life-history parameters of Culicoides (Avaritia) imicola Kieffer in the laboratory at different rearing temperatures

This laboratory study investigates the sub-adult developmental cycle of field collected Culicoides (Avaritia) imicola Kieffer (Diptera; Ceratopogonidae). The period required from blood-feeding field-collected females to the production of progeny adults occupied 34–56 days at 20 °C, 15–21 days at 25 °C and 11–16 days at 28 °C, demonstrating clear temperature dependence. When reared at 28 °C, C. imicola demonstrated higher variability in fecundity (between 2.4 and 20.6 eggs/female) and lower hatching rates (50.0–62.2%), although larval survival rates to pupation were low at all temperatures (20–30%). Similarly, the mean emergence rate from pupae was the highest at lower temperatures. These results highlight the difficulty in establishing and maintaining a laboratory colony of this species from field-collected material and results are discussed in reference to future research directions that may aid this process.