An unknown horse disease appeared in Mraclin 80 years ago - what caused “Mraclin horse disease“?

U rujnu i listopadu 1938. godine pojavila se pojedinačno u turopoljskim selima Rakarje, Buševec i Kurilovec, a kao ograničena epizootija i u Mraclinu akutna neurološka bolest konja. Bolest je dobila naziv “mraclinska bolest konja” (MBK). Od ove bolesti oboljelo je i liječeno 200 od ukupno 442 konja u selu, a 25 ih je uginulo. Svi su imali gotovo jednake akutne neurološke simptome. Uzrok bolesti nikada nije dokazan. Velika sličnost kliničkih znakova MBK s bolešću konja prouzročene virusom Zapadnog Nila (VZN) ponukali su na pretpostavku da je VZN izazvao MBK 1938. godine. Analize razudbenih i patohistoloških nalaza uginulih konja ukazuju na masivnu nekrozu jetre prouzročenu intoksikacijom, kao osnovnim uzrokom uginuća. Živčani simptomi su posljedica hepatične encefalopatije, a krvarenja i žutica su očite posljedice jetrene disfunkcije. Može se zaključiti da je jedan od mogućih uzroka mraclinske bolesti konja intoksikacija. Ovakav je zaključak utemeljen na pronađenim patološkim nalazima iz 1938. godine u arhivi Zavoda za patologiju Veterinarskog fakulteta Sveučilišta u Zagrebu. Moguće pitanje je isto tako, čime su se konji otrovali? Diferencijalno dijagnostički je teško, ali ne i nemoguće isključiti virusne uzročnike bolesti poput VZN, što bi svakako jako pomoglo u točnom otkrivanju uzroka MBK iz 1938. godine.An acute neurological disease of horses appeared sporadically in September and October of 1938 in the villages of Rakarje, Buševec and Kurilovec in the Turopolje area, and as a limited epizootic event in Mraclin. The disease was named “Mraclin Horse Disease” (MHD). Of the 442 horses in the village, 200 came down with clinical signs and were treated, and 25 perished. All the affected horses had virtually identical acute neurological symptoms. The cause of the disease has not yet been proven, but the great similarity between the clinical signs of MHD with encephalomyelitis caused by the West Nile virus (WNV), suggested that WNV may have caused MHD in 1938. However, analysis of the pathological and pathohistological findings of horses that perished from MHD indicated massive necrosis of the liver caused by unknown toxin as the main cause of death. Neurological symptoms are a consequence of hepatic encephalopathy, and bleeding and icterus are obvious consequences of liver dysfunction. It can therefore be concluded that one of the possible causes of death was intoxication. This conclusion is based on the pathological findings of 1938, found in the archives of the Department of Pathology, Faculty of Veterinary Medicine, University of Zagreb. Another question that arises is what caused the poisoning of these horses? Differential diagnosis is difficult, but it is not impossible to exclude viral pathogens such as WNV, which would certainly help to detect the causative agent of MHD in 1938

Otitis externa in dogs – microbial pathogens and antimicrobial susceptibility

Upala vanjskog slušnog kanala ili otitis externa (OE) česta je pojava u pasa. Bolest je multifaktorijalne etiologije, a primarni uzroci najčešće su: alergije, poremećaji u izlučivanju žlijezda, poremećaji epitelizacije, tumori ili polipi slušnog kanala, strana tijela u slušnom kanalu te autoimune bolesti. Infekcije bakterijama i kvasnicama predstavljaju sekundarne uzročnike upale kod OE, koji nastupaju po djelovanju primarnih uzroka i faktora koji mijenjaju mikrookoliš slušnog kanala. Predisponirajući faktori mogu biti suženja slušnog kanala, pretjerana vlažnost, endokrinopatije ili traume, a upala će uvijek biti rezultat kombinacije gore navedenih čimbenika. Najčešći patogeni mikroorganizmi koji kompliciraju upalu u pasa su stafilokoki i streptokoki od Gram-pozitivnih bakterija te bakterije roda Pseudomonas, Proteus i Escherichia od Gram-negativnih bakterija. Uz navedene bakterijske uzročnike u visokom postotku je zastupljena kvasnica Malassezia pachydermatis koja kao oportunist naseljava kožu psa. U istraživanju smo pretražili 44 obriska vanjskog slušnog kanala pasa s izraženom kliničkom slikom upale. Iz navedenih obrisaka izdvojili smo patogene bakterije i kvasnice u 25 slučajeva ili 56,81%. Najzastupljeniji patogeni bili su Staphylococcus pseudintermedius (n=8 ili 32%) i Malassezia pachydermatis (n=5, 20%), a zatim koagulaza- negativni stafilokoki (16%), streptokoki (12%), Pseudomonas aeruginosa i Bacillus spp. (8%) te Proteus mirabilis (4%). Osjetljivost izolata prema antimikrobnim lijekovima određivali smo disk difuzijskom metodom, a testirano je ukupno 14 komercijalno dostupnih antimikrobnih pripravaka. Najbolju osjetljivost izolati su pokazali prema amoksicilinu s klavulanskom kiselinom te prema gentamicinu, enrofloksacinu i cefaleksinu, a najveća rezistencija zabilježena je prema ceftiofuru, neomicinu i sulfametoksazolu s trimetoprimom. Amoksicilin s klavulanskom kiselinom pokazao je 100% učinkovitost prema Staphylococcus pseudintermedius, ali i potpunu rezistenciju kod izdvojenih Gram- negativnih bakterija. U slučajevima infekcije s Gram-negativnim bakterijama Pseudomonas aeruginosa i Proteus mirabilis, jedino su na gentamicin pokazale punu osjetljivost, dok su na ostale testirane pripravke pokazali potpunu rezistenciju. Visoki postotak bakteriološki negativnih rezultata (43,18%) najvjerojatnije je posljedica uzimanja uzoraka nakon neučinkovite terapije.Inflammation of the external ear canal or otitis externa (OE) is common in dogs. Disease is of a multifactorial aetiology, and the most common primary causes are allergies, glandular disorders, epithelial disorders, tumours or polyps of the auditory canal, foreign bodies in the auditory canal and autoimmune diseases. Bacterial and yeast infections are secondary causes of inflammation in the OE, which occur due to the action of primary causes and factors that alter the microenvironment of the auditory canal. Predisposing factors may be obstruction of the auditory canal, excessive humidity, endocrinopathies or trauma. Inflammation will always be the result of a combination of the above-mentioned factors. The most common pathogens that complicate inflammation in dogs are staphylococci and streptococci among the Gram positive bacteria, and the genera Pseudomonas, Proteus and Escherichia among the Gram negative bacteria. In addition to the above-mentioned bacterial pathogens, Malassezia pachydermatis is frequest, and settles on the skin of the dog as an opportunist. In this study, we examined 44 swabs of the external ear canal of dogs with a pronounced clinical picture of inflammation. Of these samples, pathogenic bacteria and yeasts were identified in 25 cases or 56.81%. The most common pathogens were Staphylococcus pseudintermedius (n=8 or 32%) and Malassezia pachydermatis (n=5, 20%), followed by coagulase-negative staphylococci (16%), streptococci (12%), Pseudomonas aeruginosa (8%), Bacillus spp. (8%) and Proteus mirabilis (4%). The susceptibility of isolates to antimicrobial drugs was determined by the disc diffusion method, and 14 commercially available antimicrobial drugs were tested. Bacterial isolates showed the highest susceptibility to amoxicillin with clavulanic acid (88.9%), gentamicin, enrofloxacin and cephalexin. The highest resistance of isolates was observed against ceftiofur (100%), cefovecin, neomycin and sulfamethoxazole with trimethoprim. Staphylococcus pseudintermedius strains showed 100% susceptibility to amoxicillin with clavulanic acid, unlike isolates of Gram negative bacteria which showed complete resistance to amoxicillin with clavulanic acid. In cases of infection with Pseudomonas aeruginosa and Proteus mirabilis, isolates showed full sensitivity only to gentamicin and complete resistance to other tested antimicrobials. The high percentage of negative results (43.18%) is likely the result of sampling after commenced or completed antimicrobial therapy

Nutritional specificity of protein and fat in mare milk compared to human and cow milk

Zadnjih se desetljeća kobilje mlijeko istražuje kao funkcionalna i ljekovita hrana. Sastav kobiljeg, humanog i kravljeg mlijeka znatno se razlikuje, a varira i o genetskim, fiziološkim i prehrambenim čimbenicima, kao i o uvjetima okoliša. Uspoređujući ga s humanim i kravljim mlijekom, kobilje mlijeko ima nižu energetsku vrijednost i niže vrijednosti masti. Sadržaj proteina sirutke i kazeina u kobiljem mlijeku sličan je onom u humanom za prehranu ljudi i čini ga pogodnijim od kravljega. Kobilje i humano mlijeko formiraju precipitat koji je lakše probavljiv od koaguluma kravljeg mlijeka. Vanjska membrana masnih globula, kao i raspodjela di- i tri- glicerida iz kobiljeg i humanog mlijeka su slične. Postotak nezasićenih masnih kiselina u kobiljem i humanom mlijeku veći je nego u kravljem mlijeku, a najveći dio otpada na polinezasićene masne kiseline sa srednjim i većim brojem ugljikovih atoma. Kobilje mlijeko zauzima sve više prostora u nutricionizmu zbog svojih specifičnih kemijskih i nutritivnih karakteristika.The composition of mare, human and cow milk differs significantly depending on the genetic, physiological and nutritional factors, and environmental conditions. Compared to human and cow milk, mare milk has lower energy and fat values, while the protein content of mare milk is similar to human milk. The content of whey protein and casein in mare milk makes it more suitable than cow milk for human consumption. Mare and human milk form a precipitate, which is much easier to digest than cow milk coagulum, and the outer membrane of fat globules and the distribution of di- and triglycerides from mare and human milk are also similar. The percentage of unsaturated fatty acids in mare and human milk is higher than in cow milk, most of which are polyunsaturated with a moderate to higher number of carbon atoms. Due to its specific chemical and nutritional properties, mare milk is in the focus of interest as a functional and healthy food

Antimicrobial resistance of virulent intestinal Escherichia coli in pre- and post-weaning piglets

Antimikrobna rezistencija bakterija rastu- ći je problem u svijetu, do kojeg je došlo zbog pretjerane i često nepotrebne uporabe antibiotika. Uzgoj svinja nije izuzet od spomenutog problema, a cilj ovog istraživanja bio je prika- zati rezistenciju bakterije Escherichia coli, kao jednog od najčešćih bakterijskih patogena u prasadi. U ovom istraživanju pretražena su 44 soja E. coli, izdvojena iz prasadi prije i poslije odbića, podrijetlom iz konvencionalnih uzgoja sa 6 gospodarstava Koprivničko-križevačke županije. Patogenost sojeva potvrđena je lan- čanom reakcijom polimeraze, dokazivanjem jednog ili više gena za čimbenike virulencije intestinalne E. coli. Disk difuzijskom meto- dom istražena je rezistencija E. coli prema 10 različitih antimikrobnih pripravaka. Rezultati su pokazali najmanju rezistenciju testiranih sojeva E. coli na amoksicilin s klavulanskom kiselinom (9,1 %), dok je rezistencija prema florfenikolu iznosila 40,9 % te enrofloksacinu 50 %. Prema ostalim antimikrobnim priprav- cima ustanovljena je značajna rezistencija, koja se kreće od 66,7 % prema gentamicinu i cefaleksinu, 73,3 % prema kolistinu, 80 % prema oksitetraciklinu, 86,2 % prema flumekvi- nu pa sve do 91,3 % prema sulfametoksazolu s trimetoprimom i 91,4 % prema linkomicinu sa spektinomicinom. Testirani sojevi pokazali su rezistenciju od 50 % ili više na 8 od 10 te- stiranih antimikrobnih lijekova, a 75 % pretra- ženih rezistentno je na 4 i više antimikrobnih lijekova. Rezultati ukazuju na viši postotak rezistentnih sojeva u usporedbi s dosadaš- njim istraživanjima u konvencionalnim uzgo- jima svinja, a visoka rezistencija zabilježena je i prema aminoglikozidima (gentamicin) i fluorkinolonima (enrofloksacin), što do sada nije bio slučaj. Testiranim izolatima E. coli iz- dvojenim iz prasadi prije i poslije odbića za- jednička je visoka osjetljivost na amoksicilin s klavulanskom kiselinom i visoka rezisten- cija prema linkomicinu sa spektinomicinom, sulfametoksazolu s trimetoprimom, oksite- traciklinu i flumekvinu. Sojevi izdvojeni iz prasadi prije odbića osjetljiviji su na fluorfe- nikol i enrofoksacin, a oni izdvojeni iz odbite prasadi na cefaleksin i kolistin. Prema našim saznanjima, ovo je prvo istraživanje antimikrobne rezistencije bakterije E. coli izdvojene iz prasadi, podrijetlom s malih farmi u Republici Hrvatskoj.Antimicrobial resistance in bacteria is a growing problem worldwide, due to the overuse and often unnecessary use of antibiotics. Pig farming is no exception, and the aim of this study was to demonstrate antimicrobial resistance of Escherichia coli as one of the most common bacterial pathogens in piglets. In this study, 44 strains of E. coli were isolated from pre- and post-weaning piglets, originating from conventional breeding on six holdings in Koprivnica-Križevci County, Croatia. In order to confirm the pathogenicity of strains, E. coli isolates were tested by PCR for the presence of genes for one or more virulence factors of intestinal E. coli. The disk diffusion test was used to investigate the resistance of E. coli to 10 different antimicrobial drugs. The results showed the lowest resistance of tested E. coli strains to amoxicillin-clavulanic acid (9.1%), while resistance to florfenicol was 40.9% and to enrofloxacin 50%. Other antimicrobial drugs showed significant resistance, ranging from 66.7% to gentamicin and cephalexin, 73.3% to colistin, 80% to oxytetracycline, 86.2% to flumequine, 91.3% to sulfamethoxazole-trimethoprim, and 91.4% to lincomicin-spectinomicin. The tested strains showed 50% or higher antimicrobial resistance to 8 of 10 antimicrobial drugs, and 75% of the strains were found to be resistant to four or more antimicrobial drugs. The results indicate a higher percentage of resistant strains compared to previous studies in conventional pig breeding, and a higher resistance observed for aminoglycosides (gentamicin) and fluoroquinolones (enrofloxacin), which was not previously recorded. The tested isolates of E. coli from pre- and post-weaning piglets share a high susceptibility to amoxicillin- clavulanic acid and high resistance to lincomicin-spectinomicin, sulfamethoxazole- trimethoprim, oxytetracycline and flumequin. The strains originated from pre-weaning piglets are more susceptible to florfenicol and enrofloxacin, while isolates from post- weaning piglets were more susceptible to cephalexin and colistin. This is the first study on antimicrobial resistance of pathogen intestinal E. coli isolates in piglets from small farms in the Republic of Croatia

Significance of accreditation in laboratory operations

Akreditacija laboratorija formalni je dokaz osposobljenosti za obavljanje poslova iz područja ispitivanja, prema međunarodno priznatim standardima. Unatoč povećanim početnim financijskim ulaganjima, stečena akreditacija laboratorija daje povjerenje u rezultate pretraživanja, povećava konkurentnost laboratorija na tržištu, a ujedno je i jedan od uvjeta za dobivanje statusa Službenih laboratorija u Republici Hrvatskoj. Najobjektivniji pokazatelji kvalitete rada su rezultati provedenih međulaboratorijskih ispitivanja. Važan pokazatelj su i povratne informacije koje laboratoriji dobivaju od svojih kupaca. Podizanjem razine tehničke osposobljenosti laboratorija obzirom na opremu i uvjete okoline, a posebno na edukaciju osoblja, dosegnuti su standardi suvremene laboratorijske dijagnostike i analitike.Laboratory accreditation is formal proof of competence for testing laboratories and compliance with internationally recognized standards. Despite the increased initial financial investment in the laboratory, accreditation gives confidence in its results and increases its competitiveness on the market. It is also one of the criteria required for obtaining the status of an official laboratory in Croatia. The most objective quality indicators are the results of interlaboratory trials. Another important indicator is customer satisfaction. By raising the level of technical competence of the laboratory in terms of equipment and environmental conditions, and staff training in particular, standards of modern laboratory diagnostics and analytical standards have been met

Microbiological quality of mare’s milk and trends in chemical composition by comparison of different analytical methods

In this study the quality of Croatian coldblooded mare’s milk during six months lactation period was investigated. Samples of milk were collected throughout six month’s lactation, from 22 mares and included three consecutive lactations. Physicochemical properties (pH, °SH, density) and chemical composition of raw mare milk have been studied. Fat, lactose, protein and solid non fat contents were analysed by infrared spectrophotometry and by conventional methods. Somatic cell count and microbiological parametres such as the total bacterial count, Enterobacteriaceae, coagulase positive staphylococci as well as presence of pathogens, Salmonella species and Listeria monocytogenes were also researched. Results are presented as comparison of standard and instrumental methods for chemical analysis (fat, protein, solid non fat, lactose). The mean values are presented as trends during lactation. Results were analysed by Stata 10.0. For results obtained by using different methods there were significant differences between methods for milk fat on 10th day, lactose content on 10th and 60th day, and total solids non fat on 60th day of lactation. Values of milk fat, protein, lactose and solids non fat obtained on the 40th, 60th, 120th and 180th days of lactation by IR spectrometry were compared with the value obtained on the 10th day of lactation by the same method (IR spectrometry). Milk of the Croatian coldblooded mares showed stabile chemical composition for all ingredients except lactose (p=0.0001), and high microbiological quality throughout the lactation period