Limaetal.IrishVeterinaryJournal2012,65:1 http://www.irishvetjournal.org/content/65/1/1 Iris Tréidliachta Éireann RESEARCH Open Access Glycaemia as a sign of the viability of the foetuses in the last days of gestation in dairy goats with pregnancy toxaemia Miguel S Lima1*, Rita A Pascoal2 and George T Stilwell1,3 Abstract Pregnancy toxaemia is one of the most common diseases affecting small ruminants in the last month of gestation. Nearly 80% of the foetal growth occurs in the last 6 weeks of gestation. Fat goats and goats carrying twins and triplets are at greater risk. Pregnancy toxaemia is characterized by metabolic acidosis, hypoglycaemia and ketonaemia and a very high mortality rate. In our study five does with pregnancy toxaemia showed a marked hyperglycaemia (12.4 ± 5.4 mmol/L). Although our findings are based on a small population sample (10 goats), we nonetheless postulate that hyperglycaemia could be explained by the death of the foetuses. Caesarian surgery was performed on four of the five does with hyperglycaemia (HG does). In the fifth, kidding was induced. In this group, two does had two dead foetuses, two had three dead foetuses and one does had four foetuses, only one of which was alive. Caesarian surgery was performed on all five does with hypoglycaemia (LG does). Four does of the LG group had three foetuses and one had two foetuses, all alive. The HG doe had lower rectal temperatures, lower sodium and higher urea nitrogen (BUN) in the blood when compared with the LG does. As the condition of affected does may deteriorate quickly, the results of the present study suggest that in the last days of pregnancy goats with pregnancy toxaemia and concurrent hypoglycaemia should be considered for caesarian surgery. Keywords: hyperglycemia, pregnancy toxemia, goats Background because foetal death removed the suppressing effect of The importance of glucose in the pregnant goat (and the foetus on hepatic gluconeogenesis. Smith and Sher- ewe) as the major source of energy to the foetus(es) is man [5] referred to the existence of a marked hypergly- well known. So pregnant goats are at high risk of devel- caemia in terminal cases. oping pregnancy toxaemia due to the rapid foetal The main objective of our study is to show that levels growth [1]. The energy requirements of the pregnant of glycaemia in pregnant goats with pregnancy toxaemia goat increase by a factor of 1.5 when she carries one could be an indicator of foetuses survivability. foetus and by a factor of 2 when she carries two foe- tuses [2]. Blood glucose levels in pregnant goats is gen- Results erally low, because of foetal demand. There is little The data from the physical examination of the 10 does information in literature addressing the occurrence of is presented in Table 1 and blood data is shown at hyperglycaemia in pregnant does. Bulgin [3] showed Table 2. that as the disease progresses in ewes, blood glucose When compared with the reference interval, the LG and cortisol levels may be elevated (above 3.85 mmol/L group had two does with low rectal temperature, four and 10 ng/ml, respectively) due to foetal death. Wastney does with lower heart rate, one with a high heart rate et al [4] suggested that the hyperglycaemia occurs and all five does with higher respiratory rate. In the HG group, all five does had a lower rectal temperature, one had a lower heart rate, two had a higher heart rate and *Correspondence:[email protected] 1DC,CIISA,FaculdadedeMedicinaVeterinária,UniversidadeTécnicade four had a higher respiratory rate when compared with Lisboa,PoloUniversitáriodaAjuda,Lisboa,1300-477,Portugal the reference interval. The average rectal temperature in Fulllistofauthorinformationisavailableattheendofthearticle ©2012Limaetal;licenseeBioMedCentralLtd.ThisisanOpenAccessarticledistributedunderthetermsoftheCreativeCommons AttributionLicense(http://creativecommons.org/licenses/by/2.0),whichpermitsunrestricteduse,distribution,andreproductionin anymedium,providedtheoriginalworkisproperlycited. Limaetal.IrishVeterinaryJournal2012,65:1 Page2of6 http://www.irishvetjournal.org/content/65/1/1 Table 1Data fromthe physicalexamination offive does all does. Two does, one in each group, showed swollen withpregnancy toxaemiaandhypoglycaemia (LG) and limbs (subcutaneous edema). Urine was collected from five does withpregnancy toxaemiaandhyperglycaemia two does (one LG doe and one HG doe) and analyzed (HG) with a dipstick1. Both does showed aciduria, ketonuria Parameter LG HG Referencevalues* and the HG doe had additionally glycosuria (the blood Age(years) 3.4±0.9 4.6±1.3 NA glucose of this doe was 21 mmol/L). (2-4) (3-6) The major finding in the blood data was very marked RectaltemperatureC° 37.8±1.5 34.3±1.2y 38.8-40 metabolic acidosis (low pH and HCO3-) in all 10 does. (35.3-39.1) (33-35.6) All except one LG does had Na blood levels below the Heartrate 67±36 85±33 70-90 reference range. All but one HG does had lower pCO2 (beats/min) (36-128) (44-116) blood levels. In relation to K only one HG doe had Respiratoryrate 68±39 49±21 15-30 blood levels within the reference range, all the others (breaths/min) (36-112) (28-72) were below it. Two HG does had lower Cl levels, one Rumenactivity Noactivity Noactivity 1-2 (contractions/min) had higher levels and one had levels within the reference range. In the LG group one doe had higher levels than Bodyconditionscore >4 >4 2.5-3 the reference range. Finally, all the HG does had higher Outcome Fourdead Alldead NA BUN levels while in the LG group one had higher levels Resultsareexpressedasmean±standarddeviation.Therangeofvaluesis and one had lower levels than the reference range. We showninbrackets. yp<0.005 were unable to determine the blood Cl and the BUN *PughDG[2] data in one HG doe due to a failure in the portable ana- lyzer. When the mean levels of the two groups were the HG group was significantly lower than in the LG compared, there was a statistically significant difference group (p < 0.05). All the does were recumbent, five of between the blood levels of glucose (p < 0.005), Na (p < them were able to stand and walk when forced to do so 0.005) and BUN (p < 0.05) (Table 2). (three LG does and two HG does). The other five were Following caesarian surgery in the LG group, four unable to stand or walk (two LG does and three HG were found to have three foetuses and one had two foe- does). Three does vocalized when approached (one LG tuses, all alive, in the HG group, two had two dead foe- doe and two HG does). Rumen motility was absent in tuses, two had three dead foetuses and one doe had four foetuses, only one of which was alive. Of the five does with hyperglycaemia, a caesarian sur- Table 2Blooddata collected fromfive does with gery was performed on four; the fifth, after kidding was pregnancy toxaemiaandhypoglycaemia (LG) andfive induced by injecting a combination of dexamethasone5 does withpregnancy toxaemiaandhyperglycaemia (HG) (1 mg/10 Kg BW, IM)) and dexcloprostenol6 (125 μl, Parameter LG HG Referencevalues* IM) delivered two dead fetuses. Glucose(mmol/L) 1.76±0.5 12.4±5.4y 2.75-4.13 (1.27-2.37) (6.16-21) Discussion pH 7.0±0.2 6.8±0.2 7.32-7.5 (6.80-7.26) (6.6-7.0) The blood sugar level is fairly constant in a given animal when variations, due to circadian rhythm, different pCO2(mmHg) 23.5±5.3 30.6±10.1 38-45 (16-30) (18-48) breeds, sex, nutrition and stress of handling, are taken HCO3-(mmol/L) 6.5±3.8 5.4±3.8 20-25 into account [6]. But, large differences exist between dif- (3.7-11.3) (2.7-11.9) ferent animal species. Whereas in humans and other Na(mmol/L) 140±4 131±3.4y 142-155 monogastrics, glucose levels of about 5.5 mmol/L are (135-146) (128-136) common (rising after a meal to 6.6 to 7.15 mmol/L and K(mmol/L) 2.8±0.5 3.2±0.5 3.5-6.7 falling between meals to 4.4 mmol/L), in ruminants (2.0-3.2) (2.4-3.7) blood glucose levels are usually much lower, approxi- Cl(mmol/L) 109±3.3 103±9.3 99-110 mately 2.2 to 3.3 mmol/L [6,7]. 104-113 104-113 (n=4) Regulating the blood glucose level is a complex func- BUN(mmol/L) 3.4±2.13 8.7±3.35x 1.7-3.3 tion, under the general control of the neuroendocrine 1.5-7 4.3-12.5 system, in which the liver plays a key role. This regula- (n=4) tion can be seen in Figure 1 Resultsareexpressedasmean±standarddeviation.Therangeofvaluesis The last 6 weeks of gestation in goats (and ewes) are a showninbrackets. critical period for the pregnant animal because approxi- xp<0.05 mately 80% of the foetal growth occurs during this per- yp<0.005 *PughDG(2) iod. Studies in sheep have shown that they synthesize Limaetal.IrishVeterinaryJournal2012,65:1 Page3of6 http://www.irishvetjournal.org/content/65/1/1 Figure1Regulationofthebloodglucoselevel(adaptedfrom6). about 100 g a day, but during late pregnancy this basal transporting lipoproteins out of the liver and back to rate can go up to about 180 g a day [1]. When the rate the adipose tissue [2]. of synthesis is too low, hypoglycaemia develops and the Withdrawal of large quantities of metabolites (as by animal becomes ketotic [1]. Pregnancy toxaemia in the foetus in late gestation) can overwhelm the body small ruminants occurs because of the competition for dam’s ability to mobilize metabolic substrates and this glucose between the pregnant animal and its foetuses, as leads to pronounced hypoglycaemia. Ketosis of varying these experience very rapid growth [3]. The foetus has intensity occurs associated with hypoglycaemia and developed several strategies in order to minimize the metabolic acidosis. Ketosis may result in acidosis lack of glucose that can occur in the pregnant female. because some of the hydrogen ions produced with the Ovine placenta has the ability to transfer glucose to the ketone bodies remain in the plasma, decreasing blood foetus even with a very low concentration of maternal pH [6]. blood glucose. The sheep foetus maintains low concen- Obese goats are at greater risk of developing preg- tration of plasma glucose (about 0.44 mmol/L), which nancy toxaemia, especially when they carry twins or facilitates transfer of glucose from the maternal plasma. triplets. Some authors [5,7,8] refer to this particular The sheep foetus maintains a relatively high (4.4 - 5.5 pregnancy toxaemia as “fat doe pregnancy toxaemia” mmol/L) plasma fructose concentration as a store for (estate ketosis) which is caused by over-conditioning of carbohydrate which can be utilized by its tissues. Fruc- the flock, herd or individual during early pregnancy. tose is synthesized entirely from glucose by the placenta According to Pugh [2] pregnant goats should have a and cannot readily pass through the placenta into body condition score of between 2.5 and 3 some 45 maternal circulation. Even though fructose is more days before parturition. All ten goats of this study had abundant than glucose in the plasma of foetal sheep, the a body condition scores above 4. This was assessed by foetal tissues utilize about twice as much more glucose palpation of the sternum [9] Goats are considered to than fructose. Foetal tissues have an amazing capacity to be tropical/subtropical animals and body fat stores that remove glucose from plasma at very low concentrations develop are laid down not in subcutaneous tissue but [1]. in the omentum and the mesentery and around the In all pregnant animals the foetus continuously with- kidneys. For this reason, conventional body condition draws metabolites, especially glucose and aminoacids, so scoring based on lumbar palpation can be misleading; it is logical that in small ruminants the main metabolic another approach is to utilize lumbar scoring in paral- disorders, or production disease, associated with carbo- lel with assessing a score on the same scale by palpat- hydrate, fat and protein metabolism is pregnancy toxae- ing the sternum, and assessing its degree of fat cover mia (usually in multiple pregnancies). Fatty liver is [9]. almost always associated with this and is frequently During late gestation, the abdominal space is filled found when necropsies are performed on these animals. with accumulated fat and an ever-expanding uterus. It is well known that ruminants are not efficient at Because of the lack of rumen space, these females have Limaetal.IrishVeterinaryJournal2012,65:1 Page4of6 http://www.irishvetjournal.org/content/65/1/1 difficulty consuming enough feedstuff to satisfy their According to some authors, this increase in BUN can be energy requirements [2]. caused by increased protein catabolism, by decomposing The great accumulation of fat in the abdominal cavity foetuses or by terminal kidney failure [13]. According to and a very small rumen were consistent findings Marteniuk and Herdt [14], pregnancy toxaemia is often observed during the necropsies in our study. A marked accompanied by dehydration, electrolyte abnormalities fatty liver was also observed in three necropsies carried and renal failure. Experimental models of ketonaemia out (two HG doe and one LG doe). have shown that systemic hypertension can occur in In sheep and goats, pregnancy ketosis is much more pregnant ewes after as little as 24 hours of food depriva- common in highly prolific selected breeds [5]. All the tion. Renal dysfunction begins with the onset of hyper- does in this study were from the Saanen breed. The tension and results in as much as a 51% decrease in mere fact that there were more Saanen goats (1100) on glomerular filtration, as well as being indicated by a rise this farm than Alpine goats (600) is probably not a satis- in BUN values and protein loss in the urine [3]. In our factory explanation for this. We had not (prior to this study, the levels of BUN were elevated in the HG does study) observed more abdominal accumulation of fat in when compared with the LG does (p < 0.05). Kidney Saanen goats than in Alpines. Therefore we think that failure could explain the lower levels of sodium that also the discrepancy is probably better explained by the Saa- occurred in the HG does (p < 0.005). However, urine nen breed being more likely to be pregnant with multi- samples obtained from two does did not show ple foetuses than are Alpines. proteinuria. According to Rook [8] clinical cases are typically lim- A marked hypokalemia could be observed in nine ited to older goats and ewes during their second or sub- does. In general, changes in the pH of the extracellular sequent pregnancies. In our study, the ranges of ages of fluid produces reciprocal H+ and K+ shifts between the the does were between 2 and 4 years in the LG group cells and the extracellular fluid. As a result, K+ tends to and between 3 and 6 years in the HG group. move into the cells with alkalemia and out of the cells Pregnancy toxaemia is a disease characterized by a with acidemia. These pH-induced effects, however, are high mortality rate and on the farm where this study transient and frequently overridden by concurrent varia- was done is the main cause of does deaths. In our study, tions in other mechanisms that influence K+ transport only one doe survived (10%). [15]. Hypokalemia has been described in humans with The diagnosis of pregnancy toxaemia was achieved liver failure, including the acute fatty liver during preg- from the history (last days of pregnancy) and by physical nancy [16]. In human patients with ketoacidosis and exam and was confirmed by a blood analysis. ketonuria, there is marked loss of K+ in the urine lead- Blood levels of glucose in affected animals varied dra- ing to hypokalemia [15]. matically and this gave rise to the idea that hypoglycae- Hypokalemia could be explained in part because the mia might indicate that the foetuses are alive and goats were not eating and, therefore, their dietary K hyperglycaemia that the foetuses are dead. Because not intake would have been reduced. We suggest that these many practitioners can measure glucose in the blood as two mechanisms, acting together, were the cause of the we did in our study, this information is not available in hypokalemia that was observed in this study. the field. It would be much easier if a urine sample Kidding was induced in one doe in the HG group could be collected to measure glucose concentration. using dexamethasone and prostaglandin; it is arguable Glucose is not found in the urine of normal domestic as to whether the hyperglycemia was due to the effect of animals unless the blood glucose increases above the the dexamethasone. In order to clarify this point, blood renal threshold, which is thought to be around 5.5 to was taken from 10 additional doe whose kidding was 7.7 mmol/L in ruminants [10]. However, in goats it induced in the previous 48 to 24 hours and blood glu- appears to be more difficult to collect a urine sample cose levels were determined. In these doe, mean glucose than in cows or ewes although there is evidence that levels were 3.2 ± 0.7 mmol/L. This would suggest that they usually urinate when forced to stand up [11]. corticosteroid did not have an effect in the blood glu- The withdrawal of large quantities of metabolites can cose levels in this group. A doe that is carrying 2 or 3 overwhelm the body’s ability to mobilize metabolic sub- foetuses and with a marked negative energy balance has strates, and can result in severe hypoglycaemia, variable all its gluconeogenic mechanisms working very effi- degrees of ketosis and metabolic acidosis [12]. Ketone ciently, so the administration of a corticosteroid has a bodies (beta-hydroxybutyrate and acetoacetate) are very modest effect, if any, on blood glucose levels. Glu- strong acids [1,6,12], and their accumulation in the cocorticoid blood concentrations are elevated in ovine blood leads to metabolic acidosis (ketoacidosis). The ketosis [1]. They can inhibit glucose utilization, and situation can progress to an irreversible stage, where force the animal’s body to rely more on free fatty acids there is dehydration and increased BUN values [12]. and ketone metabolism for caloric needs [1]. Prolonged Limaetal.IrishVeterinaryJournal2012,65:1 Page5of6 http://www.irishvetjournal.org/content/65/1/1 hypoglycaemia can lead to hyperactivity of the adrenal producing milk, low producing goats, dry does and glands, with increased cortisol secretion, insulin activity young goats) can be seen in Table 3. antagonism and to effective inhibition of maternal glu- Milk production in this herd averaged approximately 3 cose use [12]. L per goat per day. Machine milking was performed twice daily. In this farm there are three kidding seasons Conclusions per year, in January, April and October. Each kidding The blood levels of glucose in pregnant goats with preg- season began on the first day of the month, and contin- nancy toxaemia can be a good indicator of the viability ued for 45 days. of the foetuses. As the condition of affected does may Four vaccine products were routinely administered to deteriorate quickly, the results of the present study, the goats on this farm: [1] A commercial 7-way clostri- although based on a small sample, suggest that in the dial bacterin2 was administered to kids at 60 days of last days of pregnancy goats with pregnancy toxaemia age, and a booster dose given 30 days later. This vaccine and concurrent hypoglycaemia should be considered for is administered to adult goats and bucks twice a year caesarian surgery. This will avoid prolonged poor wel- [2]. An autogenous Corynebacterium pseudotuberculosis fare for the doe and its kids. In the future, in order to (caseous lymphadenitis) vaccine was administered to further strengthen these findings, we would like to con- kids at 60 days of age, and a booster dose given 30 days firm the viability of the foetuses by ultrasonography. later. Subsequently, the vaccine was administered annually to all yearling and adult goats [3]. An autoge- Methods nous Mannheimia haemolytica vaccine was adminis- This study was performed on farm with 1700 dairy tered to kids at 7 to 15 days of age, and a booster dose goats, 30 miles northeast of Lisbon. The goats were administered at 45 days of age. Subsequently, this same from two breeds: Saanen (1100 animals) and Alpine vaccine was administered to animals of all ages at 6- (600 animals). The goats were housed in confined straw month intervals [4]. A commercial Mycoplasma agalac- yards, had access to free stalls and were fed a complete tiae vaccine3 was administered to all adult goats, one ration, ad libidum. Their Total Mixed Ration (TMR) month before the beginning of each kidding season, the consisted of corn silage, rye-grass hay, alfalfa hay, young does receiving a booster 3 weeks later. brewer’s grain and a concentrate mixture. The feed was Twice annually, fecal samples were collected from distributed once a day. All the adult goats had free every pen on the farm, and McMaster’s fecal egg counts access to mineral blocks. One month before kidding were performed on those samples. Groups of goats were they started eating wheat straw ad libidum and 1 Kg of treated with an anti-helmintic drug, where nematode concentrate distributed 4 times a day. Nutrient analysis egg counts of 250 eggs or more per gram of feces were of the TMR for the different groups of goats (high obtained. However, during the conduct of this study, Table 3Nutrient analysis ofthe TMRforthe different groupsofgoats (high producing milk, low producing goats,dry does andyoung goats) Nutrient HighProduction LowProduction Dry Younggoats(from6monthsuntilonemonthbefore Goats Goats Does kidding) DryMatterintake(Kg/ 2.4-3 1.8-2 1.4-1.5 0.8-0.94 day) Drymatter% Max:50 Max:50 80-90 40 Forage%DM 40 40 50 CrudeProtein(%) 17-18 16-17 12 12 CrudeFat(%) 5 5 4 3 CrudeFiber(%) 16 17 17 17 AcidDetergentFiber(%) 19 20 21 21 NeutralDetergentFiber 28 30 31 32 (%) Starch(%) 15-18 12-14 10-15 10-20 Ca%DM 0.8-0.9 0.75-0.8 0.35 0.35 P%DM 0.4-0.5 0.35-0.4 0.25 0.25 Mg%DM 0.25-0.3 0.25-0.3 0.2 0.2 Salt%DM 0.4-0.5 0.4-0.5 0.25 0.25 Limaetal.IrishVeterinaryJournal2012,65:1 Page6of6 http://www.irishvetjournal.org/content/65/1/1 nematode egg counts did not exceed this threshold, and Competinginterests no goats were ever treated for internal parasites. It is Theauthorsdeclarethattheyhavenocompetinginterests. common knowledge that dairy goats maintained in dry Received:19August2011 Accepted:23January2012 lot have minimal problems with gastro-intestinal nema- Published:23January2012 todes [2]. This study involved 10 does with clinical signs of References 1. BergmanEN:DisordersofCarbohydrateandFatMetabolism.InDukes pregnancy toxaemia. The does were in two groups, a PhysiologyofDomesticAnimals..11edition.Editedby:SwensonMJ,Reece group of does with hypoglycaemia (LG group) and a WO.CornellUniversityPress; 1993:492-516. group with hyperglycaemia (HG group). A physical 2. PughDG:Diseasesofthegastrointestinalsystem.InSheepandGoat MedicineEditedby:PughDG,SaundersWB2002,69-105. exam was performed and a blood sample was taken 3. Bulgin:MSPregnancyToxemia.In-depthReview,Wool&Wattles2005, from the jugular vein and analyzed immediately at the 33:9-10. farm with a portable analyser4 for glucose, pH, pCO2, 4. 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Ethics Committee (Comissão de Ética e Bem-Estar Ani- 9. HarwoodD:Thedifferencesandsimilaritiesbetweendiseasesofsheep andgoats.GoatVeterinarySocietyJournal25:37-40. mal da Faculdade de Medicina Veterinária). 10. CarlsonGP:ClinicalChemistryTests.InLargeAnimalInternalMedicine..2 edition.Editedby:SmithBP.MosbyYearBookInc; 2006:441-469. Statistical analysis 11. EastNE:PregnancyToxemia,AbortionsandPeriparturientDiseases.Vet ClinNorthAm1983,5:601-618,LargeAnimalPractice. A two-tailed T test for independent samples was used to 12. SargisonND:Pregnancytoxaemia.InDiseasesofSheep.Editedby:Aitken compare the means values of the two groups, LG and ID.BlackwellPublishing;2007:359-362. HG [17]. 13. AndrewsA:PregnancyToxaemiaintheEwe.Practice1997,19:306-312. 14. MarteniukJV,HerdtTH:Pregnancytoxemiaandketosisofewesand does.VetClinNorthAm1988,4:307-315,FoodAnimalPractice. Endnotes 15. RoseBT,PostTW:Hypokalemia.ClinicalPhysiologyofAcid-Baseand 1. Combur10 Test, Roche ElectrolyteDisorders.5edition.McGraw-Hill,MedicalPublishingDivision; ® 2001,836-887. 2. Miloxan , Merial 16. LeeWM:AcuteLiverFailure.TheNewEnglandJournalofMedicine1993, ® 3. Agalactivax , Schering-Plough 329(N°25):1862-1872. 4. i-Stat, Sensor Devices Incorporated, Waukesha, 17. SnedecorGW,CochranWG:Thecomparisonoftwosamples.Statistical Methods.7edition.IowaUniversityPress;1980,83-106. Wisconsin, USA 5. Vetacort®, Vetoquinol doi:10.1186/2046-0481-65-1 ® Citethisarticleas:Limaetal.:Glycaemiaasasignoftheviabilityof 6. Gestavet-Prost , Hipra thefoetusesinthelastdaysofgestationindairygoatswithpregnancy toxaemia.IrishVeterinaryJournal201265:1. Acknowledgements TheauthorswouldliketothankMr.AntonioBaraoforallowingtheauthors tocarryoutthisstudyonhisfarmandtoMr.MichaelBourneforreviewing themanuscript. Theresearchleadingtotheseresultshasreceivedfundingfromthe EuropeanUnion’sSeventhFrameworkProgramme(FP7/2007-2013)under grantagreementN°266213. Authordetails Submit your next manuscript to BioMed Central 1DC,CIISA,FaculdadedeMedicinaVeterinária,UniversidadeTécnicade and take full advantage of: Lisboa,PoloUniversitáriodaAjuda,Lisboa,1300-477,Portugal.2Barãoe Barão,CoutadaVelha,Benavente,2130-010,Portugal.3AWIN-Animal • Convenient online submission WellfareIndicators,FaculdadedeMedicinaVeterinária,UniversidadeTécnica deLisboa,PoloUniversitáriodaAjuda,Lisboa,1300-477,Portugal. • Thorough peer review • No space constraints or color figure charges Authors’contributions • Immediate publication on acceptance MLdidmostofthefieldwork(physicalexaminations,bloodcollection).RP identifiedthesickgoatsandhelpedwiththefieldwork.GShelpedtodraft • Inclusion in PubMed, CAS, Scopus and Google Scholar andtorevisethemanuscript.Allauthorsreadandapprovedthefinal • Research which is freely available for redistribution manuscript. Submit your manuscript at www.biomedcentral.com/submit