Spatiotemporal analysis of African swine fever outbreaks on South African smallholder farms, 1993-2018.

African swine fever (ASF) is a contagious viral disease of swine worldwide. ASF in South Africa has for many years been confined to a controlled area in the northeast of the country that was proclaimed in 1935. Since 2012, outbreaks are more likely to occur in the historically ASF-free area. This study aimed to analyse the spatial and spatiotemporal structure of ASF outbreaks in South Africa between 1993 and 2018. Global space-time clustering of ASF outbreaks was investigated by the Diggle space-time K-function while Kulldorff's spatial scan statistic was applied to detect local cluster of ASF outbreaks. Globally, ASF outbreaks exhibit statistically significant spatial clustering. They have shown a significant negative space-time interaction at month scale (p = 0.003) but no significant space-time interaction at year scale (p = 0.577), revealing strong evidence that ASF cases that are close in space occur in months which are close and vice versa. In studying local area space-time clustering at both month and year scale, three significant local clusters associated with high-rate were detected. These clusters are localised in both the ASF-controlled area and outside the controlled area with radius varying from 60.84 km up to 271.43 km and risk ratio varying from 6.61 up to 17.70. At month scale, clusters with more outbreaks were observed between June 2017 and August 2017 and involved 22 outbreaks followed by the cluster that involved 13 outbreaks in January 2012. These results show the need to maintain high biosecurity standards on pig farms in both inside and outside the ASF-controlled areas.

Faecal Escherichia coli as biological indicator of spatial interaction between domestic pigs and wild boar (Sus scrofa) in Corsica.

On the Mediterranean island of Corsica, cohabitation between sympatric domestic pigs and Eurasian wild boar (Sus scrofa) is common and widespread and can facilitate the maintenance and dissemination of several pathogens detrimental for the pig industry or human health. In this study, we monitored a population of free-ranging domestic pigs reared in extensive conditions within a 800-ha property located in Central Corsica which was frequently visited by a sympatric population of wild boar between 2013 and 2015. We used GPS collars to assess evidence of a spatially shared environment. Subsequently, we analysed by PFGE of XbaI-restricted DNA if those populations shared faecal Escherichia coli clones that would indicate contact and compared these results with those collected in a distant (separated by at least 50 km) population of wild boar used as control. Results showed that one of eight wild boars sampled in the study area shed E. coli XbaI clones identical to clones isolated from domestic pig sounders from the farm, while wild boar populations sampled in distant parts of the study area shared no identical clone with the domestic pigs monitored. Interestingly, within the sampled pigs, two identical clones were found in 2013 and in 2015, indicating a long-time persisting colonization type. Although the method of isolation of E. coli and PFGE typing of the isolates requires intensive laboratory work, it is applicable under field conditions to monitor potential infectious contacts. It also provides evidence of exchange of microorganisms between sympatric domestic pigs and wild boar populations.

Assessing the extent and use of risk analysis methodologies in Africa, using data derived from the Performance of Veterinary Services (PVS) Pathway.

African countries that wish to export are increasingly faced with import risk assessments from importing countries concerned about the sources of their imported goods. Other risk analysis methodologies and approaches are also employed, which focus on animal and human health within countries and communities. Based on an analysis of evaluations conducted by the World Organisation for Animal Health (OIE), using the Performance of Veterinary Services Tool, the authors attempt to define current practice in Africa and degrees of compliance with the World Trade Organization Agreement on the Application of Sanitary and Phytosanitary Measures ('SPS Agreement') and OIE standards. To assist in this task, the authors also make use of a review of selected risk assessment reports. Results point to a lack of technical capacity and capability to conduct risk assessments in compliance with OIE standards (except in the case of three countries), ranging from an outright absence of any form of (documented) risk assessment and consecutive risk management decisions (level of advancement 1) to shortcomings in one or several aspects of the risk assessment process. This is confirmed by a number of case studies, half of which have been produced by international consultants. The major recommendations of this paper are i) to strengthen the human resources pool for conducting risk assessments and ii) to establish dedicated risk assessment units, with clear terms of reference, job descriptions and policies, procedures and protocols.

Les pays africains souhaitant accéder aux marchés d'exportation sont confrontés à la réalisation de plus en plus fréquente d'appréciations du risque à l'importation par les pays importateurs, qui cherchent à vérifier les sources d'approvisionnement des marchandises qu'ils importent. D'autres méthodologies d'analyse du risque sont également pratiquées, axées sur la santé animale et la santé humaine au niveau des pays et des communautés. À partir d'une analyse des évaluations réalisées par l'Organisation mondiale de la santé animale (OIE) au moyen de l'Outil d'évaluation des performances des Services vétérinaires, les auteurs tentent de définir les pratiques actuelles en Afrique ainsi que le niveau de conformité des pays à l'Accord sur l'application des mesures sanitaires et phytosanitaires de l'Organisation mondiale du commerce (« Accord SPS ¼) et aux normes de l'OIE. Pour ce faire, les auteurs s'appuient également sur l'analyse d'une sélection de rapports d'appréciation du risque. Les résultats laissent apparaître l'inadéquation des capacités et des compétences techniques dédiées à l'appréciation du risque par rapport aux normes de l'OIE (à l'exception du cas de trois pays), depuis l'absence totale d'appréciation du risque à partir d'éléments documentés et de décisions relevant de la gestion de ce risque (niveau d'avancement 1), jusqu'à des insuffisances concernant un ou plusieurs aspects du processus d'appréciation du risque. Cette analyse est confirmée par une série d'études de cas, dont la moitié a été produite par des consultants internationaux. Les auteurs recommandent principalement de renforcer les ressources humaines dédiées aux appréciation du risque et de mettre en place des unités dédiées, en déterminant clairement leur mandat, leur fonction, ainsi que les politiques à mener, les procédures applicables et les protocoles à suivre.

Cada vez más, los países africanos que desean exportar son sometidos a determinaciones del riesgo de importación por parte de los países importadores, preocupados por la procedencia de las mercancías que importan. A veces también se emplean otros métodos y planteamientos de análisis del riesgo, centrados en determinados aspectos zoosanitarios y sanitarios de los países o comunidades de origen. Basándose en un análisis de las evaluaciones realizadas por la Organización Mundial de Sanidad Animal (OIE) con la Herramienta PVS, los autores tratan de aprehender los usos actualmente vigentes en África, así como los niveles de cumplimiento del Acuerdo sobre la Aplicación de Medidas Sanitarias y Fitosanitarias (Acuerdo MSF) de la Organización Mundial del Comercio (OMC) y las normas de la OIE, ayudándose también para ello del estudio de una serie de informes de determinación de riesgos. Los resultados ponen de relieve la falta de competencias y capacidades técnicas para llevar a cabo determinaciones de riesgos con arreglo a las normas de la OIE (salvo en el caso de tres países), con situaciones que van desde la ausencia absoluta de toda forma de determinación (documentada) de los riesgos y, a partir de ahí, de toda decisión para gestionarlos (nivel de progreso 1), hasta la existencia de uno o varios aspectos defectuosos en el proceso de determinación de riesgos, observación que confirman una serie de estudios monográficos, la mitad de ellos obra de consultores internacionales. Las principales recomendaciones de los autores apuntan a fortalecer la dotación de recursos humanos para efectuar determinaciones de riesgos y a establecer unidades dedicadas específicamente a esta labor, habiendo definido con toda claridad su mandato, los perfiles que deben integrarlas y sus políticas, procedimientos y protocolos de trabajo.

Experimental Evaluation of Faecal Escherichia coli and Hepatitis E Virus as Biological Indicators of Contacts Between Domestic Pigs and Eurasian Wild Boar.

Domestic pigs and Eurasian wild boar (Sus scrofa) share several important viral and bacterial pathogens. Therefore, direct and indirect contacts between domestic pigs and wild boar present a risk of pathogen spillover and can lead to long-term perpetuation of infection. Biological indicators could be a powerful tool to understand and characterize contacts between wild boar and domestic pigs. Here, faecal Escherichia coli and Hepatitis E virus (HEV) were explored as potential biological indicators under experimental conditions. The data gained in our pilot study suggest that faecal E. coli can be used as biological indicator of contact between wild boar and domestic pig. For HEV, faecal transmission was also confirmed. However, molecular studies on full-genome basis did not reveal markers that would allow tracing of transmission direction. Based on these promising results, future field studies will especially target the practicability of E. coli microbiome molecular typing as surrogate of contacts at the wildlife-livestock interface.

Living at the edge of an interface area in Zimbabwe: cattle owners, commodity chain and health workers' awareness, perceptions and practices on zoonoses.

BACKGROUND: In the great Limpopo transfrontier conservation area (GLTFCA), there is an increased interface between wildlife and domestic animals, because rural households move their cattle into the game park in search of grazing and watering resources. This creates opportunities for inter-species transmission of infectious diseases, including zoonoses like brucellosis and tuberculosis, which may also pose a health risk to the local rural communities. This study investigated the awareness, perceptions and practices on zoonoses amongst rural cattle owners, commodity chain- and health-workers in three different localities around Gonarezhou National Park (GNP), Zimbabwe, where the interface between wild and domestic animals varies. METHODS: A cross-sectional study was conducted in Malipati, Chikombedzi and Chiredzi that are considered to be high-, medium- and low-domestic animal-wildlife interface areas, respectively. Data was collected from cattle owners, commodity chain and health-workers using a semi-structured questionnaire. To determine the public health risk of food-borne zoonoses, their practices with regard to meat and milk consumptions, and measures they take to prevent exposure to infections were assessed. Data were analyzed using descriptive statistics and principal component analysis. RESULTS: Most respondents (52.8 %, 102/193) were cattle owners, followed by health (30.1 %, 58/193) and lastly commodity chain workers (17.1 %, 33/193). Overall 67.4 % (130/193) of the respondents were aware of zoonoses with respective 48, 81.8, and 93.1 % of cattle owners, commodity chain, and health workers, being aware. Significantly more cattle owners (P < 0.05) from medium and low interface areas were aware of zoonoses compared to those from high interface areas. All categories of respondents cited anthrax (69.2 %), rabies (57.7 %), tuberculosis (41.5 %) and brucellosis (23.9 %) as important zoonoses. About half (46.1 %; 89/193) of the respondents perceive wildlife as important reservoirs of zoonoses. High proportions 98.4 % (190/193) and 96.4 % (186/193) of the respondents indicated that they consume meat and milk, respectively. Access to game meat and milk from informal markets was closely associated with consumption of raw meat and milk. CONCLUSIONS: Fewer cattle owners from a high interface area of Malipati are aware of zoonoses compared to other areas due to combined effects of limited education and other factors disadvantaging these marginalised areas. This may increase their risk of exposure to zoonoses, considering that consumption of raw meat and milk is common. Thus, awareness campaigns may reduce the public health impact of zoonoses at the interface.

Epidemiological surveillance methods for vector-borne diseases.

Compared with many other diseases, the ever-increasing threat of vector-borne diseases (VBDs) represents a great challenge to public and animal health managers. Complex life cycles, changing distribution ranges, a variety of potential vectors and hosts, and the possible role of reservoirs make surveillance for VBDs a grave concern in a changing environment with increasing economic constraints. Surveillance activities may have various specific objectives and may focus on clinical disease, pathogens, vectors, hosts and/or reservoirs, but ultimately such activities should improve our ability to predict, prevent and/or control the diseases concerned. This paper briefly reviews existing and newly developed tools for the surveillance of VBDs. A range of examples, by no means exhaustive, illustrates that VBD surveillance usually involves a combination of methods to achieve its aims, and is best accomplished when these techniques are adapted to the specific environment and constraints of the region. More so than any other diseases, VBDs respect no administrative boundaries; in addition, animal, human and commodity movements are increasing dramatically, with illegal or unknown movements difficult to quantify. Vector-borne disease surveillance therefore becomes a serious issue for local and national organisations and is being conducted more and more at the regional and international level through multidisciplinary networks. With economic and logistical constraints, tools for optimising and evaluating the performance of surveillance systems are essential and examples of recent developments in this area are included. The continuous development of mapping, analytical and modelling tools provides us with an enhanced ability to interpret, visualise and communicate surveillance results. This review also demonstrates the importance of the link between surveillance and research, with interactions and benefits in both directions.

Multivariate analysis of traditional pig management practices and their potential impact on the spread of infectious diseases in Corsica.

Corsica is a French Mediterranean island with traditional extensive pig farming oriented towards the production of high quality cured meat products. The increasing success of these cured products in continental Europe has triggered the development and organisation of an extensive pig farming industry. However, these pig farming practices have seldom been described and analysed to understand the potential risk of introduction and spread of infectious diseases. We conducted a cross-sectional study in Corsica in 2013 to characterise the main pig management practices and to identify groups of farms with similar practices and therefore homogeneous risk of introduction and spread of infectious diseases. We interviewed 68 pig farmers and investigated different farm management practices which could lead to contact between herds, such as trading animals, sharing pastures, feed and reproduction management (direct contacts), slaughtering and carcass waste management, and contacts with people and vehicles (indirect contacts). The practices were described and the farms grouped by multiple factor and hierarchical clustering analyses. Results revealed interesting patterns in the introduction and spread of infectious disease, such as the seasonality of pig production, the potential local spread of diseases in pastures due to the presence of free-ranging boars, carcasses, and animal waste. Multivariate analyses identified four groups of farms with different levels of risk of the spread of infectious disease, illustrating changes in farmers' customs from free-range uncontrolled farming systems to more controlled systems aimed at the production of high quality pork products. These results will be useful to more realistically simulate the spread of infectious diseases among Corsican pig farms and highlight the need for awareness raising campaigns among the stakeholders to reduce risky practices.

Bridge hosts for avian influenza viruses at the wildlife/domestic interface: an eco-epidemiological framework implemented in southern Africa.

Wild terrestrial birds can act as potential local spreaders or bridge hosts for avian influenza viruses (AIVs) between waterfowl (the maintenance hosts of AIVs) and domestic avian populations in which AIVs may cause disease. Few studies have investigated this hypothesis, although it is an important knowledge gap in our understanding of AIV spread within socio-ecosystems. We designed a simple and reproducible approach in an agro-ecosystem in Zimbabwe based on: (1) bird counts at key target sites (i.e., wetlands, villages, intensive poultry production buildings and ostrich farms) to identify which wild birds species co-occur in these different sites and seasons when the risk of AIV transmission through these potential bridge hosts is maximal and (2) targeted sampling and testing for AIV infection in the identified potential bridge hosts. We found that 12 wild bird species represented the vast majority (79%) of co-occurrences in the different sites, whereas 230 bird species were recorded in this ecosystem. Specifically, three species - barn swallow, Hirundo rustica, red-billed quelea, Quelea quelea and cattle egret, Bubulcus ibis - represented the main potential bridge host species (65% of co-occurrences). In two out of these three species (i.e., barn swallow and red-billed quelea), we detected AIV infections, confirming that they can play a bridge function between waterfowl and domestic species in the ecosystem. Our approach can be easily implemented in other ecosystems to identify potential bridge hosts, and our results have implications in terms of surveillance, risk management and control of AIV spread in socio-ecosystems.

Preliminary assessment of bovine tuberculosis at the livestock/wildlife interface in two protected areas of northern Botswana.

Protected areas of northern Botswana such as the Okavango Delta (OD) or Chobe National Park (CNP) are well-known hot spots for the conservation of African wildlife. However, their infection status regarding bovine tuberculosis (BTB) at the domestic/wildlife interface has never been investigated. To provide preliminary baseline data on the circulation of Mycobacterium bovis in those sites, we performed a cross-sectional survey on 130 buffalo in both protected areas (60 individuals from CNP and 70 from OD) and 818 cattle in their surrounding communal lands (369 in CNP and 449 in the OD). Whole-blood samples were tested using a commercial interferon-gamma assay (IFN-γ) with modifications. The apparent BTB prevalence in buffalo was nil in CNP and 0.7% 95% CI [0.2-1.9] in the OD, while the apparent BTB prevalence in cattle was 0.7% 95% CI [0.2-2.1] in the OD and 2.4% 95% CI [1.2-4.7] in CNP. True prevalence values calculated on the basis of the locally applicable IFN-γ test performance suggested that BTB prevalence was nil in both buffalo populations and in cattle from the OD interface, but reached 2.3% 95% CI [0.2-4.5] in cattle populations around CNP. The results of a questionnaire survey conducted among a sample of farmers living in the communities adjacent to each conservation area (97 and 38 persons in the OD and CNP, respectively) suggested a higher risk of the circulation of M. bovis at the wildlife/livestock interface of the CNP than at that of the OD. However, further comprehensive studies are needed to confirm the circulation of M. bovis and to monitor the inter-species and transboundary transmission of BTB in northern Botswana.

Outbreaks of African horse sickness in Senegal, and methods of control of the 2007 epidemic.

Since first being detected in Nigeria in January 2007, African horse sickness virus serotype 2 (AHSV-2) has spread throughout the northern hemisphere, and was first reported in Senegal. A retrospective study was conducted from December 2009 to April 2010 using data collected in the field combined with information available at the Direction of Veterinary Services. The epidemic started in the Dakar region with two outbreaks in March and June 2007, respectively, and spread in several parts of the country between July and November 2007. During this period, 232 outbreaks and 1137 horse deaths were reported. The epidemic was controlled by mass vaccination using a polyvalent-attenuated vaccine. This retrospective study was conducted with various assumptions of AHSV-2 introduction, and provides recommendations for implementing an early warning surveillance system for African horse sickness in Senegal.

Review of the sylvatic cycle of African swine fever in sub-Saharan Africa and the Indian ocean.

African swine fever (ASF) is a major limiting factor for pig production in most of the countries in Sub-Saharan Africa and the Indian Ocean. In the absence of vaccine, a good understanding of the ecology and epidemiology of the disease is fundamental to implement effective control measures. In selected countries of Southern and East Africa, the association between Ornithodoros moubata ticks and warthogs has been described in detail in the literature. However, for many other countries in the region, information related to the sylvatic cycle is lacking or incomplete. In West African countries, for instance, the role of wild pigs in the epidemiology of ASF has never been demonstrated and the existence and potential impact of a sylvatic cycle involving an association between soft ticks and warthogs is questionable. In other countries, other wild pig species such as the bushpigs (Potamochoerus spp.) can also be asymptomatically infected by the virus but their role in the epidemiology of the disease is unclear and might differ according to geographic regions. In addition, the methods and techniques required to study the role of wild hosts in ASF virus (ASFV) epidemiology and ecology are very specific and differ from the more traditional methods to study domestic pigs or other tick species. The aim of this review is (i) to provide a descriptive list of the methodologies implemented to study the role of wild hosts in African swine fever, (ii) to compile the available knowledge about the sylvatic cycle of ASFV in different regions of Sub-Saharan Africa and the Indian Ocean in addition to the one that has been described for East and Southern Africa, and (iii) to discuss current methodologies and available knowledge in order to identify new orientations for further field and experimental surveys.

The mosquito Aedes (Aedimorphus) vexans arabiensis as a probable vector bridging the West Nile virus between birds and horses in Barkedji (Ferlo, Senegal).

Active catches of adult females of Aedes vexans arabiensis Patton, (Diptera: Culicidae) Patton by nets or aspirator, were conducted in 2003 and 2004 in the vegetation at the edge of temporary ponds in Barkedji, Senegalese Ferlo area. Two hundred and forty-one engorged females were captured, dissected and the gut content adsorbed on a Whatman filter paper and analysed using the enzyme-linked immunosorbent assay (ELISA) technique to determinate the bloodmeal origin. Results indicated that Ae v. arabiensis fed primarily on mammals, including horses (35.7% of the bloodmeals), but also on birds (10%). Moreover, associations between horses and birds accounted for 42% of the mixed bloodmeals. These results show an opportunistic feeding behaviour and suggest that Ae v. arabiensis is a probable vector bridging the West Nile virus between horses and birds hosts in the Ferlo area.

Environmental risk factors of West Nile virus infection of horses in the Senegal River basin.

In 2005, a serological study was carried out on horses in five ecologically contrasted zones of the Senegal River basin (Senegal) to assess West Nile virus (WNV) transmission and investigate underlying environmental risk factors. In each study zone, horses were randomly selected and blood samples taken. A land-cover map of the five study areas was built using two satellite ETM+ images. Blood samples were screened by ELISA for anti-WNV IgM and IgG and positive samples were confirmed by seroneutralization. Environmental data were analysed using a principal components analysis. The overall IgG seroprevalence rate was 85% (n=367; 95% CI 0.81-0.89). The proximity to sea water, flooded banks and salted mudflats were identified as protective factors. These environmental components are unfavourable to the presence of Culex mosquitoes suggesting that in Senegal, the distribution of the vector species is more limiting for WNV transmission than for the hosts' distribution.

Activation of myosin light chain phosphatase in intact arterial smooth muscle during nitric oxide-induced relaxation.

We investigated whether myosin light chain phosphatase activity changes during nitric oxide-induced relaxation of contracted intact carotid media and how changes in phosphatase activity mediate this relaxation. We also investigated one mechanism for regulating this phosphatase. Myosin phosphatase activity, myosin light chain phosphorylation, guanosine 3',5'-cyclic monophosphate (cGMP) concentration, and phosphorylation of the inhibitory protein CPI-17 were all assayed in homogenates of one carotid media ring at each time point during nitric oxide-induced relaxation. The application of sodium nitroprusside to histamine-contracted media caused rapid declines in light chain phosphorylation and force. These were temporally correlated with a rapid elevation of cGMP and a large transient increase in myosin phosphatase activity. During the early response to nitroprusside, when force declined, increases in myosin phosphatase activity, concurrent with cGMP-mediated decreases in calcium and myosin light chain kinase activity, could accelerate light chain dephosphorylation. CPI-17 was dephosphorylated upon application of nitroprusside at the same time that myosin phosphatase activity increased, suggesting that the removal of inhibition by phospho-CPI-17 contributed to the increase in myosin phosphatase activity. After 20 min of nitroprusside, myosin phosphatase activity had declined to basal levels, however low force was sustained. Additional light chain phosphorylation-independent mechanisms may be involved in sustaining the relaxation.

Relationship of Ca2+ sparks to STOCs studied with 2D and 3D imaging in feline oesophageal smooth muscle cells.

We recorded Ca2+ sparks and spontaneous transient outward currents (STOCs) simultaneously in smooth muscle cells using whole-cell patch recording and a unique, high-speed widefield digital imaging system to monitor fluo-3 fluorescence in both two and three dimensions (2D and 3D). In 2D imaging, the correlation between the amplitude of a spark and its corresponding STOC was a weak one, and 27 % of the sparks failed to cause STOCs. The STOCless sparks were not significantly different in amplitude from those that caused STOCs. Three-dimensional imaging disclosed that STOCless sparks were located close to the cell surface, and on average their apparent distance from the cell surface was not significantly different from the sparks that cause STOCs. Statistical evaluation of spark clusters disclosed that there were regions of the cell where the probability of spark occurrence was high and others where it was quite low.

Susceptibility of high and low producer dairy goats to single experimental infection with Trichostrongylus colubriformis.

Culled dry dairy goats, which differed in their level of production in previous lactations, received a single infection with Trichostrongylus colubriformis. The objectives of the study were twofold. First, the study aimed at examining the ability of dairy goats to develop an immune response to trichostrongyle infection and the associated cellular changes developing within the intestinal mucosa. Second, a comparison between animals differing in their level of production was assessed, in order to determine whether these differed in their susceptibility to infection. No difference occurred in egg excretion, worm burden and local inflammatory cell responses between high (HP) and low (LP) producer dairy goats, in contrast to observations in previous studies. Because their nutrition was controlled and milk production absent in the goats employed in the present study, these results suggest that any genetic component associated with the selection of HP and LP goats had little influence on the development of acquired resistance to T. colubriformis. The analysis of the relationship between different cell types in the mucosa and some characteristics of the worm population show that eosinophils are negatively related to worm burden. Also, a role is suggested for mast cells and globule leukocytes in the modulation of egg excretion.

A questionnaire survey on the practices adopted to control gastrointestinal nematode parasitism in dairy goat farms in France.

To evaluate the efficacy with which recommendations of means to avoid the spread of anthelmintic resistance in parasitic nematode populations are communicated to farmers on dairy goat farms in France, a questionnaire survey was undertaken on their use of antiparasitic drugs. Information was collected from 73 farms in two main areas of dairy goat production. The data referred to three years. Anthelmintics were used in 69 farms, the mean number of treatments per year being 2.74. Changing the drug from one year to another was not practised. Moreover, of the 58 farms using two or more treatments per year, only 37% used anthelmintics from different classes in the lactation and drying-off periods. Benzimidazoles and probenzimidazoles were given in all except two farms and these substances represented more than 80% of all the treatments. Levamisole/pyrantel or avermectins were used in 15% and 27% of the farms, respectively. Double the ovine dose, as recommended in goats to ensure efficacy of benzimidazoles, was applied in 55% of the farms. In addition, in all the flocks, the substances were given on the basis of a mean estimated live weight and not by reference to the heaviest animal. These results indicate that errors in the use of anthelmintics are still frequent in dairy goat farms in France, with probable consequences for the spread of anthelmintic resistance in the populations of parasites.

Efficacy of copper oxide needles for the control of nematode parasites in dairy goats.

The spread of benzimidazole-resistant nematodes in dairy goat farms is of a great concern as probably more than 70% of the flocks are involved. As there are very few other anthelmintic options during the lactating period, we have evaluated the efficacy of copper oxide needles (CON, Copinox, Bayer, UK) in both experimental and natural infections in goats. The curative effect of CON (2-4 g) on existing worm burdens was assessed in goats experimentally infected with Teladorsagia circumcincta, Haemonchus contortus and Trichostrongyus colubriformis, compared to controls. The preventive effect of CON (4 g) on worm establishment was monitored for 2 months in animals experimentally infected with H. contortus and for 3 months in naturally infected animals on a farm exhibiting predominant infections with T. circumcincta and Oesophagostomum venulosum. In both experimental and natural conditions, the efficacy of CON was nil against Teladorsagia, Trichostrongylus and Oesophagostomum infections. In contrast, the efficacy of CON against Haemonchus was clearly established in reducing the worm burden (75%) as well as in lowering the egg output (37-95%) in relation to the establishment of new infections over several weeks. Copper oxide needles may represent an alternative to conventional anthelmintics in the control of Haemonchus infection in some goat farms.

A dilution immunoassay to measure myosin regulatory light chain phosphorylation.

We examined the quantitation of myosin regulatory light chain phosphorylation (MRLCP) by Western blot and found both offset and saturation errors. The desirable characteristics of an MRLCP assay are that the dynamic range be 60- to 100-fold and that the detection threshold be known and preferably very small relative to total MRLC concentration. No technique examined provided all these characteristics. However, accurate measurements can be obtained by including serial dilutions of the sample to provide a fractional calibration scale in terms of the dephosphorylated light chain and by using interpolation of the phosphorylated band signal intensity to provide values for the relative phosphorylation ratio. We found that this method offers several advantages over methods that rely on signal ratios from single samples: The dilution ratio method is less subject to errors from differences in protein load, it offers estimates of the error in the individual measurement, and has some redundancy that increases the likelihood of obtaining a valid measurement despite gel or membrane artifacts.