Comparison of detection procedures of Mycoplasma hyopneumoniae, Mycoplasma hyosynoviae, and Mycoplasma hyorhinis in lungs, tonsils, and synovial fluid of slaughtered pigs and their distributions in Thailand

The aim of this study was to investigate whether direct PCR (DP) gave similar results to culture prior to PCR (CPP) for detecting mycoplasmas in different types of pig tissues. A total of 724 samples obtained from lungs, tonsils, or synovial fluids from 270 slaughtered pigs were used. The history of clinical signs, lung score, and the presence of joint lesions were recorded during sample collection. The rates of detection of Mycoplasma hyopneumoniae, Mycoplasma hyosynoviae, and Mycoplasma hyorhinis using both procedures were evaluated. The overall prevalences of M. hyopneumoniae, M. hyosynoviae, and M. hyorhinis were 40.3%, 12.3%, and 64.6%, respectively, and the detection rate depended on the sample type and the procedure used. With lung tissue, DP gave a higher detection rate for M. hyopneumoniae (77.4%) than CPP (38.5%). M. hyorhinis was detected by CPP at 15.6% and 18.1% and by DP at 31.5% and 5.2%, respectively. The positive rate derived from tonsil from CPP was closed to that of DP. Using synovial fluid could not yield any positive M. hyorhinis from CPP whereas 37.2% was positive from DP. In contrast, using sample tissue from lung and tonsil by CPP could show much higher positive number than that of DP. There was a significant relationship between joint lesion and M. hyorhinis detection by DP (P < 0.05) but not for M. hyosynoviae and M. hyorhinis detected by CPP. We speculated that lung was a proper sample for M. hyopneumoniae and M. hyorhinis detection by DP and CPP, respectively. Tonsil was likely the community of persistent M. hyosynoviae and M. hyorhinis with highly detection by CPP. Synovial fluid was apparently unsuitable for mycoplasmal culture. The accuracy of mycoplasmal detection may depend upon the type of sample relevant to the detection procedure used.     

Cloning and expression analysis of PRL and PRLR genes in black Muscovy duck

ABSTRACT

1. Prolactin hormone (governed by the PRL gene) is secreted by the anterior pituitary of animals, which combines with its receptor (prolactin receptor, PRLR) to act on target cells. Both PRL and PRLR are mainly associated with reproductive performance. The genetic mechanism of nesting in poultry is not yet clear, and so the aim of the current study was to determine expression patterns of PRL and PRLR at different times across the breeding stages of black Muscovy ducks.

2. In this study, the CDS regions of PRL and PRLR were determined by RACE sequencing. The expression levels of PRL and PRLR in the pituitary, ovary and uterus from the black Muscovy duck were compared and analysed during the pre-laying, laying and nesting periods.

3. The results showed that PRL and PRLR are highly homologous in a variety of poultry species. The expression of the PRL gene in the pituitary was the highest, which was significantly higher than seen in the ovary and uterus. This trend ran through the entire prenatal period, i.e. the laying period and the nesting period. The expression level of the PRLR gene in the pituitary and ovary was generally low, and expression in the uterus was the highest. There was no significant difference in expression of the PRLR gene between pituitary and ovary during different periods, but the expression level of the PRLR gene in the uterus reached its highest level during the nesting stage, which was significantly higher than seen in the early laying period.     

Embryo transfer,a useful technique to be applied in small community farms?

The objective of this study was to determine if the technique of embryo transfer in cattle can be commercially feasible in a region situated in the humid tropics of Mexico. Twenty-six Bos taurus and twenty-six Bos indicus cows were estrous synchronized and superovulated to obtain a total of 80 embryos of both sub-species. Embryos were classified using stereoscopic microscopy based on established criteria. Nine dual-purpose farms situated in the tropics of Mexico were chosen to provide ten recipient cows each to transfer one embryo per cow. The females were transferred using a fixed-time protocol after verifying the presence of a corpus luteum on the seventh day after the end of hormonal treatment. Pregnancy diagnosis was carried out 28 days after embryo transfer by ultrasonography. Estimation of the cost was determined by calculating the expenses for preparation of the donor and embryo recovery, which were US 633 and US633 and US 589 for B. taurus and B. indicus, respectively. The cost of each embryo was determined considering the number of transferable embryos recovered, which was 3.8 on mean. The cost of each conception was calculated taking into account the percentage of pregnant animals (27% on mean), and the cost for preparing donor and recipient cows, for transferring embryo. The overall cost per gestation was US 1,447. Considering a 50:50 ratio of male to female born, the cost for a replacement heifer calf was US1,447. Considering a 50:50 ratio of male to female born, the cost for a replacement heifer calf was US 2,894, which surpassed by far the commercial cost of a crossbred ready-to-bred heifer normally used as replacement (approximately US $900).     

Genetic relatedness of methicillin-resistant Staphylococcus pseudintermedius (MRSP) isolated from a dog and the dog owner

In the present study four methicillin-resistant Staphylococcus pseudintermedius (MRSP) strains isolated from a dog (n = 3) and the anterior nares of the dog owner (n = 1) were investigated by conventional and molecular methods. The species identity of the four S. pseudintermedius strains was confirmed by conventional methods, by PCR mediated amplification of S. intermedius/S. pseudintermedius specific segments of thermonuclease encoding gene nuc and by restriction fragment length polymorphism analysis of phosphoacetyltransferase encoding gene pta. Investigation of the four S. pseudintermedius for toxinogenic potential revealed that all four strains were positive for the exfoliative toxin encoding gene siet and the leukotoxin encoding genes lukS, lukF. The oxacillin and penicillin resistance of the four S. pseudintermedius strains could be determined by cultivation of the strains on oxacillin resistant screening agar base, ChromID MRSA Agar and Brilliance MRSA Agar and by multiplex PCR detecting the resistance genes mecA and blaZ. The genetic relatedness of the strains was studied by macrorestriction analysis of their chromosomal DNA using pulsed field gel electrophoresis (PFGE). According to PFGE all four S. pseudintermedius strains represent an identical bacterial clone indicating a cross transmission between the dog and the dog owner.     

Number of nitrate- and nitrite-reducing Selenomonas ruminantium in the rumen, and possible factors affecting its growth

The cell number of Selenomonas ruminantium (S. ruminantium) that reduces nitrate and nitrite in the rumen was usually 8–10% of the total number of S. ruminantium (an order of 10⁶/mL). The percentage was not affected by the roughage/concentrate ratio or nitrate content of the diet in 2 weeks. However, feeding a high‐nitrate diet for 12 weeks increased the percentage. The percentage of lactate‐using S. ruminantium, such as the ssp. lactilytica, was less than 1% of the total number of S. ruminantium. No S. ruminantium was found that used formate as an electron donor for nitrate and nitrite reduction. Lactate and H₂ appeared to be important for nitrate and nitrite reduction by S. ruminantium. Nitrate reduction by S. ruminantium was enhanced by the coexistence of amylolytic bacteria in a medium containing starch, and as a result, nitrite accumulation increased. Coexistence of cellulolytic bacteria facilitated the growth of S. ruminantium in a medium containing cellulose, and consequently increased nitrite reduction. In order to suppress nitrite accumulation in the rumen, it may be important to enhance fiber digestion.     

Nutritive value of some herbage for dromedary camels in the central arid zone of Iran

The purpose of this study is to prepare standard tables of the chemical composition of feedstuff and to determine the digestibility and palatability of different plant species in the dromedary camel, this research was conducted considering the consumed herbages by camels in the central arid zone of Iran. The following plant species were included: Alhagi camelorum, Artemisia sieberi, Atriplex lentiformis, Haloxylon persicum, Hammada salicornica, Salsola tomentosa, Salsola rigida, Seidlitzia rosmarinus, Suaeda fruticosa, Tamarix tree, and Tamarix kotschi. Thirty samples of the browsing parts were collected from three sites in the rangelands of Qom and Yazd province. The chemical composition of the samples, including dry matter, crude protein (CP), crude fiber, neutral detergent fiber (NDF), acid detergent fiber (ADF), ether extract, total ash, macroelements (Ca, P, Mg, K), microelements, and gross energy were measured. The in vitro digestibility of the plants was measured by camel liquor using the Tilley and Terry method. The palatability of the plants was measured by four mature camels in cafeteria trials. Data were analyzed by general linear model method using the SAS software. The highest CP (17.5%) related to Haloxylon persicum and the lowest NDF (26.2%) and ADF (12.6%) were related to Salsola rigida. The lowest CP (5.5%) and the highest NDF (72.8%) and ADF (59.6%) were related to Artemisia sieberi. The results also indicate that Atriplex lentiformis, Alhagi camelorum, Seidlitzia rosmarinus, Suaeda fruticosa, Haloxylon persicum, Salsola tomentosa, Hammada salicornica, T. kotschi, Salsola rigida, Tamarix tree, and Artemisia seiberi were more pleasurable feeds, respectively. There was no consistent relationship between the palatability of herbages with the percentage of digestible organic matter in the dry matter or chemical composition.     

An aroA Mutant of Edwardsiella ictaluri Is Safe and Efficacious as a Live,Attenuated Vaccine

Abstract

The aroA gene of Edwardsiella ictaluri was cloned and sequenced, and the sequence data were used to construct a deletion–insertion mutation in the aroA gene. The mutated gene was transferred into a virulent, wild-type E. ictaluri strain by conjugation and allelic exchange. Putative aroA mutants were confirmed phenotypically by demonstrating a need for supplementation with aromatic metabolites to support growth in minimal media. The genetic construction was evaluated by using the polymerase chain reaction to amplify appropriate regions of the aroA deletion–insertion, and DNA sequencing of the amplified products confirmed the predicted construction. A selected mutant, LSU-E1, was passed 30 times in nonselective media with no reversion to the wild-type following screening of 1.6 × 10¹¹ colony-forming units. The mutant was demonstrated via injection to be attenuated more than 5 logs₁₀ compared with the wild-type E. ictaluri strain, and it was avirulent by immersion and oral routes. Tissue persistence studies indicated that the mutant maintained the ability to invade following immersion exposure, but no viable cells were detected after 48–72 h. Significant levels of protection from disease were demonstrated following immersion vaccination of channel catfish Ictalurus punctatus.     

Effects of recombinant interferon-gamma on cytokine secretion from monocyte-derived macrophages infected with Salmonella typhi

Salmonella typhi (S. typhi) is an important pathogen which causes typhoid fever. The cytokines released from the macrophages, playing a role in the host defense against Salmonella infection, are crucial in the defense against the infection. IFN-γ provides a protection against Salmonella infection by developing macrophage activation in different mechanisms. This study was designed to investigate the effect of the recombinant IFN-γ (rIFN-γ) on the cytokines secreted from S. typhi stimulated macrophages. Macrophage isolation was done in the heparinized blood samples obtained from healthy people, and following the priming with rIFN-γ for 72 h the cells were stimulated by S. typhi and then the cytokine levels in culture supernatants were determined by enzyme-linked immunosorbent assay. It was observed that rIFN-γ reversely increased the levels of IL-1, IL-2 the levels of which were decreased by S. typhi and that it increased TNF-α levels while suppressing the levels of antiinflammatory cytokines such as IL-10 and TGF-β the levels of which were increased by S. typhi. Consequently, rIFN was observed to increase protective Th1 response by affecting the secretion of cytokine during S. typhi infection and it was considered to be a good target especially to prevent and treat invasive Salmonella infections.     

ermB-mediated erythromycin resistance in Streptococcus uberis from bovine mastitis

The ermB gene was identified in 111 erythromycin resistant isolates of Streptococcus uberis from cases of bovine mastitis associated either with a constitutive (47/111) or an inducible (64/111) phenotype, as well as a phenotypic resistance to all macrolides tested. Resistance to lincosamides was identified in 14 other isolates of S. uberis from bovine mastitis cases and was mainly mediated by the linB gene; resistance conferred by a combination of two genes (linB–lnuD, ermB–linB) was also detected.     

Effect of cyclodextrin diallyl maleate on methane production, ruminal fermentation and microbes in vitro and in vivo

Effects of β‐cyclodextrin diallyl maleate (CD‐M) on methane production, ruminal fermentation and digestibility were studied both in vitro and in vivo. In in vitro study, diluted ruminal fluid (30 mL) was incubated anaerobically at 38°C for 6 and 24 h with or without CD‐M using hay plus concentrate (1.5:1) as a substrate. The CD‐M was added at different concentrations (0, 1.25, 2.5, 5.0 and 7.5 g/L). The pH of the medium and numbers of protozoa were not affected by the addition of CD‐M. Total volatile fatty acids were increased and ammonia‐N was decreased, molar proportion of acetate was decreased and propionate was increased (P < 0.05) by CD‐M. Methane was inhibited (P < 0.05) by 14–76%. The effect of CD‐M on methane production and ruminal fermentation was further investigated in vivo using four Holstein steers in a cross‐over design. The steers were fed Sudangrass hay and concentrate mixture (1.5:1) with or without CD‐M (2% of feed dry matter) as a supplement. Ruminal proportion of acetate tended to decrease and that of propionate was increased (P < 0.05) 2 h after CD‐M dosing. Total viable counts, cellulolytic, sulfate reducing, acetogenic bacteria and protozoa were unaffected while methanogenic bacteria were decreased (P < 0.05) by CD‐M. The plasma concentration of glucose was increased, whereas that of urea‐N was decreased (P < 0.05). Methane was inhibited (P < 0.05) from 36.4 to 30.1 L/kg dry matter intake by the addition of CD‐M. Apparent digestibilities of dry matter and neutral detergent fiber were not affected while that of crude protein was increased (P < 0.05) in the medicated steers. These data suggested that dietary supplementation of CD‐M decreased methane production and improved nutrient use.     

An investigation into the distribution, host-preference and population density of Ixodid ticks affecting domestic animals in Bangladesh

To study the distribution, host-preference and population density of ixodid ticks in Bangladesh, an attempt was made to collect adult ticks from various host animals in three distinct topographic zones, viz. flood plains, hills and steppe ‘Barind’. Five species of ixodid ticks were recorded, namely, Boophilus microplus (56.3%), Haemaphysalis bispinosa (11.3%), Rhipicephalus sanguineus (14.7%), Hyalomma anatolicum anatolicum (15.0%) and Amblyomma testudinarium (2.8%). The data showed that B. microplus occurred predominantly on cattle (42.4%). The other hosts involved were buffaloes (12.5%), goats (25.5%) and pigs (8.2%). H. bispinosa mostly parasitized goats (31.5%) rather than cattle (12.0%) and buffaloes (10.8%). R. sanguineus was principally a dog tick (27.4%) but also parasitized cattle (10.8%) and goats (6.8%). H. a. anatolicum was restricted to cattle (19.2%) and A. testudinarium was found on both cattle (4.4%) and pigs (2.3%). These results indicate that ixodid ticks are not strictly host-specific except for H. a. anatolicum. The population density of these ticks was significantly (p < 0.01) influenced by the changing of seasons. B. microplus, H. bispinosa and R. sanguineus were by far the most widely distributed species; the distribution of H. a. anatolicum was restricted to the steppe ‘Barind tract’ and A. testudinarium was found in the hilly regions only.     

Detection of superantigenic toxin genes in Staphylococcus aureus strains from subclinical bovine mastitis

The aim of this study was to determine the presence of genes encoding enterotoxins (sea-sej) and toxic shock syndrome toxin-1 (tst) of Staphylococcus aureus strains (n = 130) isolated from subclinical bovine mastitis in Turkey by polymerase chain reaction. Sixty-one (46.9%) isolates were found to contain one or more toxin genes. The most frequently found enterotoxin genes were seg (16.2%) and sei (16.2%), followed by sec (15.4%), sed (10.8%), and sej (10.8%), respectively. The tst gene was detected in seven (5.4%) isolates. None of S. aureus strains harbored sea, seb, see, and seh genes. Since these toxins are recognized agents of staphylococcal food poisoning, it must be considered that the consuming raw milk and raw milk products would pose public health risk as high prevalence of toxigenic S. aureus was found in this study.     

Characterization of Francisella tularensis Strains,Comparing Their Carbon Source Utilization

Thirteen Francisella tularensis strains were isolated from 22 seropositive brown hares (Lepus europaeus) originating from different parts of Hungary, and further two from a patas monkey (Erythrocebus patas) and vervet monkey (Chlorocebus aethiops). The isolates were identified as F. tularensis ssp. holarctica on the basis of culture, morphological and biochemical characteristics. The identification was verified by polymerase chain reaction and the sequencing of the partial 16S rRNA gene. Utilization of carbon sources of the 15 F. tularensis strains was characterized with the Biolog system. The system was able to identify the strains already after 4 h of incubation, not only after the standard 24 h. After the analysis and comparison of the metabolic profiles of our strains with the Biolog database, it was concluded that not all carbon sources indicated in the database were utilized by our isolates. The Biolog software fails to distinguish the highly virulent F. tularensis ssp. tularensis and the moderately virulent F. tularensis ssp. holarctica but the Biolog microplates can be manually read to differentiate the two subspecies based on glycerol source utilization. As all the studied strains were unable to use glycerol, they could be identified as F. tularensis ssp. holarctica. The dendrogram based on the metabolic relationship of the strains shows that the isolates are very similar to each other, which correlates with the conservative genetic character of F. tularensis ssp. holarctica.     

Lentivirus-mediated RNA interference of myostatin gene affects MyoD and Myf5 gene expression in duck embryonic myoblasts

1. The aim of this study was to construct lentivirus-mediated short hairpin RNA (shRNA) vectors targeting the duck MSTN gene and investigate whether these vectors can affect the development of duck primary cultured embryonic myoblasts.

2. MSTN mRNA levels in the myoblasts were detected using quantitative real-time polymerase chain reaction (PCR), cell proliferation was assessed by MTT assays and cell differentiation was assayed by photography.

3. MSTN mRNA levels in PLL3.7-MSTN-shRNA1, PLL3.7-MSTN-shRNA2 and PLL3.7-MSTN-shRNA3 lentivirus-mediated shRNA groups were reduced by 61.6%, 76.9% and 79.1%, respectively, compared to control cells.

4. Down-regulation of MSTN in duck embryonic myoblasts stimulated cell proliferation and inhibited differentiation, accompanied by a greater than twofold down-regulation of MyoD expression and up-regulation of Myf5 expression.

5. These results revealed that silencing of MSTN changes the development of duck embryonic myoblasts by regulating the expression level of MyoD and Myf5 genes.

     

动物源性沙门氏菌的耐药性分析及氟苯尼考类耐药基因的鉴定

为了解动物沙门氏菌的流行情况和药物敏感性及氟苯尼考耐药株的耐药基因分布,本试验对临床上疑似患沙门氏菌病的病料进行病原分离和细菌的多重PCR鉴定;采用K-B法测定分离株对23种抗菌药物的敏感性;选择氟苯尼考耐药菌株扩增floR、fexA、fexB、cfr和pexA基因。结果显示,共鉴定出61株沙门氏菌,其中肠炎沙门氏菌10株,鸡白痢沙门氏菌12株,鼠伤寒沙门氏菌39株。所有菌株对青霉素、红霉素、万古霉素耐药,90.16%对6种及6种以上抗菌药耐药。floR基因广泛存在于鼠伤寒沙门氏菌氟苯尼考耐药菌株中(8/12,66.67%),未发现其他耐药基因。研究结果表明鼠伤寒沙门氏菌是鹅源分离株中的优势血清型;floR基因主要介导沙门氏菌对氟苯尼考耐药性,但可能还存在其他机制。     

Anti‐microbial Susceptibility of Streptococcus spp. Isolated from Bovine Mastitis in Argentina

The in vitro susceptibility to penicillin G, erythromycin and clindamycin was determined by the disc diffusion test and by E‐test for a total of 47 streptococcal strains (three Streptococcus uberis, 36 Streptococcus agalactiae, eight Streptococcus dysgalactiae spp. dysgalactiae) isolated from bovine intramammary infections in Argentina. Moreover, resistance phenotypes of erythromycin‐resistant streptococcal isolates was characterized. MIC₉₀ of penicillin G, erythromycin and clindamycin for S. agalactiae were 0.75, 8.0 and 12.0 μg/ml respectively. Resistance to erythromycin and clindamycin was detected in 13 (27.6%) and 12 (25.5%) isolates respectively. No isolate was resistant to penicillin G. Resistance against macrolides, lincosamides and streptogramin B (MLS_B) represented by the constitutive MLS_B phenotype was present in 11 (23.4%) erythromycin‐resistant isolates and two isolates (4.3%) expressed the M phenotype. The inducible MLS_B phenotype was not identified. Results suggest that beta‐lactams are the first‐line antibiotics when treating streptococcal udder infections; however, the continuous monitoring of the antibiotic resistance is essential, as the emergence of resistant strains has become a growing concern on the therapy of bovine mastitis.     

Evaluation of the grazing potential of grass species in Tarchonanthus veld of the northern cape 1

Abstract

The grazing value of the seven most prominent grass species in Tarchonanthus‐limeveld was investigated from 1977 to 1981.

The frequency of utilization of Chrysopogon serrulatus was exceptionally high (93%), while Sporobolus fimbriatus, Themeda triandra and Digitaria seriata were intermediate (50–60%) followed by Cymbopogon plurinodis (34%). Eragrotis lehmanniana and Fingerhuthia africana were poorly grazed (12 to 16%). The utilization of species was influenced by stocking rate. Statistical differences between species indicated a relatively high biomass production (600 to 740 g DM/m² per year) for E. lehmanniana, C. plurinodis and T. triandra, while F. africana, S. fimbriatus and D. seriata were intermediate (320 to 420 g DM/m² per year). E. lehmanniana, C. plurinodis and T. triandra produced the highest above‐ground phytomass, but had a lower crude protein content (2,9 to 7,4%) and digestibility (46,1 to 61,3%) than the other four species (3,8 to 9,7% crude protein and 50,5 to 68,4% digestibility).     

Biochemical Characterization of Mycoplasma bovirhinis,Mycoplasma dispar and Recent Bovine Isolates of Mycoplasma canis

The pattern and kinetics of substrate utilization by the type strains of Mycoplasma canis, M. bovirhinis and M. dispar and ten recent M. canis isolates from cattle were determined. Metabolism of a range of sugars and organic acids by M. dispar was detectable by measurement of oxygen uptake. Organic acids were not utilized by M. bovirhinis or M. canis, and there was no oxygen uptake during metabolism of glucose or other sugars, as monitored by a pH-change method. The M. canis strains varied in their ability to metabolize sugars; seven of the isolates from cattle had the distinctive ability to metabolize sucrose, and one isolate, plus the type strain (from a dog), metabolized N-acetylglucosamine. The M. bovirhinis strain metabolized maltose. However, all the test strains oxidized glycerol at high rates and with a high affinity. Oxidation of glycerol has been reported for other mycoplasmas from the bovine respiratory tract and leads to the production of hydrogen peroxide, a potential virulence factor.     

Intra-articular stabilisation of the equine cricoarytenoid joint

Reasons for performing study: The success of laryngoplasty is limited by abduction loss in the early post operative period. Objective: To determine the efficacy of polymethylmethacrylate (PMMA) in stabilising the cricoarytenoid joint (CAJ) and reducing the force on the laryngoplasty suture. Hypothesis: Injection into the cricoarytenoid joint resists the forces produced by physiological laryngeal air flows and pressures thereby reducing the force experienced by the laryngoplasty suture. Methods: Ten cadaver larynges were collected at necropsy and PMMA was injected into one CAJ at selected random. Each larynx was subjected to physiological conditions with with constant (static) or cycling (dynamic) flow. The specimens were tested sequentially in each of 4 conditions: 1) bilateral full abduction (Control 1); 2) transection of the suture on the side without PMMA; 3) bilateral abduction achieved by replacing the suture (Control 2); and 4) cutting the suture on the PMMA side. Tracheal pressure and flow and pressure in the flow chamber were recorded using pressure and flow transducers. The strain experienced by each suture during bilateral abduction (Controls 1 and 2) was measured. Statistical comparison of the 4 conditions was performed using a mixed effect model with Tukey's post hoc test for multiple comparisons. The strain gauge data were analysed by paired comparison of the regression slopes. Results: In the static and dynamic states, tracheal pressure increased and tracheal flow decreased when the suture on the non‐cement side was cut (P<0.05). There was no significant difference in any outcome measure between PMMA injected into the CAJ and bilaterally abducted specimens (Controls 1 and 2) for either condition. The rate of increase in strain with increasing translaryngeal pressure was significantly less on the suture with PMMA placed in the CAJ (P = 0.03). Conclusions: These data provide strong evidence that injecting PMMA into the CAJ resists the collapsing effect of physiological airflows and pressures in vitro and reduces the force experienced by the laryngoplasty suture during maximal abduction. Potential relevance: Augmentation of prosthetic laryngoplasty with this technique may reduce arytenoid abduction loss in the early post operative period.     

Fur animal botulism hazard due to feed

To assess the botulism hazard in fur animal feed production, 236 fur animal feed components and feed samples were analysed for Clostridium botulinum by detecting BoNT-encoding genes (botA, botB, botC, botE or botF) by PCR and for sulphite-reducing clostridia (SRC) by iron sulphite agar. The quality of the hazard analysis of critical control points (HACCP) –based in-house control system (IHCS) was evaluated with respect to botulism risk in feed plants (n = 32). The overall prevalence of C. botulinum was 13% in different feed components and 5% in feed. The estimated MPN count of C. botulinum in feed components was 6.4 × 10³/kg at the highest and was shown to poorly correlate with SRC count. The critical control points in IHCSs were variable, and control limits were improperly set in most feed-producing plants. C. botulinum possesses a persistent safety hazard for fur animals by feed production, and control practices should be reassessed.