Development of villus atrophy in the small intestine of sheep infected with Trichostrongylus rugatus

Pairs of sheep infected with 120 000 larvae of Trichostrongylus rugatus were killed at intervals from 2 to 56 days after infection (DAI). Worms were located in tunnels in the epithelium of villi or upper intestinal crypts at all stages of development. Villus atrophy developed progressively until 16 DAI, when surface microtopography, characterised by subtotal villus atrophy, stabilised. Most severe lesions were in the first 3 m of small intestine where the density of nematodes was highest. Discontinuities in the epithelium and effusion of inflammatory cells and tissue fluids into the lumen were rare. No effects of infection on body-weight gain, appetite or serum total protein and albumin were evident in the first 20 days of infection. However, it was concluded that T. rugatus fundamentally resembled T. colubriformis and T. vitrinus in the response it elicited in the intestine of sheep, and must be considered potentially pathogenic.     

Dual infection of lambs with Trichostrongylus colubriformis and chlamydiae

Lambs in one group were inoculated with chlamydiae and Trichostrongylus colubriformis larvae simultaneously; lambs in a second group were inoculated with chlamydiae after a T. colubriformis infection was established. Results were compared to those from groups inoculated with T. colubriformis only and chlamydiae only. There were no significant differences in fecundity associated with chlamydial inoculation. Packed cell volumes (PCVs) were depressed in groups inoculated with chlamydiae, and chlamydial antibody formation appeared to be inhibited when chlamydiae was superimposed on a T. colubriformis infection. Relatively few T. colubriformis adults were recovered in lambs inoculated with chlamydiae and T. colubriformis simultaneously.     

Resistance dynamics of calves to infection with Trichostrongylus axei

Calves were immunized with 10 000 infective larvae each of Haemonchus contortus, Ostertagia ostertagi, and Trichostrongylus axei and challenged with 100 000 of each species. When compared with nonimmunized controls, the immunized calves had 95% fewer H. contortus, 10% fewer O. ostertagi, and 20% fewer T. axei; the latter two reductions were not significant. In four other experiments single or multiple immunizing inoculations of T. axei followed by challenge at various intervals after immunization failed to result in significant resistance measured by the number of worms of the challenge inoculation that became established, by post-challenge fecal egg count, and by gross pathologic alterations of abomasa at necropsy.     

Pathological changes in the small intestine of neonatal calves naturally infected with reo-like virus (rotavirus).

Nine calves, of which six had been challenged with an enteropathogenic strain of Escherichia coli, were found to be naturally infected with rotavirus. Rotavirus was recovered from the faeces of six calves and rotavirus antigen was detected in the intestinal mucosa of all calves. Stunting and fusion of villi were seen principally in the proximal and middle small intestine, where rotavirus antigen was detected by immunofluorescence. Typical lesions of enteric colibacillosis were found in the distal ileum of the challenged calves, associated with adhesion of the challenge strain of E coli to the mucosa. All samples were removed from the intestine under general anaesthesia and denudation of villi was not observed. However, following exsanguination and resampling at the same site in the small intestine of one calf, denudation was a constant feature.     

The response of sheep vaccinated with irradiated Trichostrongylus colubriformis larvae to impulse and sequential challenge with normal larvae

Sheep aged 9–10 months were vaccinated with three doses of 20 000 irradiated Trichostrongylus colubriformis larvae at fortnightly intervals. This gave 97–99% protection against subsequent “impulse” challenge with 40 000 normal larvae or “sequential” challenge with 2 000 larvae/week-day for 4 weeks.The sheep were protected against both types of challenge when worms from the immunizing infections were present at the time of challenge, and when these worms were first removed by anthelmintic treatment.     

Ultrastructural changes in the small intestine of neonatal calves with enteric colibacillosis

The small intestines of calves inoculated orally with the enteropathogenic strain of Escherichia coli 0101:K'B41',K99 were examined by electron microscopy at 3, 6, 12, 16, 21, 36, 69, 70 and 72 hours after inoculation. The challenge organism adhered to the mucosa of the distal small intestine from six hours post-inoculation. Bacteria were separated from the microvillous brush border by a gap of 200 to 300 nm in which bacterial fimbriae and the microvillous glycocalyx were seen. Bacteria never were found in epithelial cells but were present in macrophages in the lamina propria from 12 hours. At three and six hours, cytopathic changes were not seen in the small intestine, but from 12 hours epithelial cells on affected villi had blunt and thick microvilli and contained cytoplasmic inclusions. Epithelial cells were seen frequently in the process of extrusion from the villi, either singly, in small groups, or as ribbons of cells. Intervillous bridges, characteristic of villous fusion, were seen frequently from 69 hours.     

The effect of γ-radiation on the development of infective larvae of Trichostrongylus colubriformis in guinea pigs and sheep

Infection of guinea pigs with normal Trichostrongylus colubriformis larvae was characterized by an initial logarithmic decline of total worms recovered over the first few days of infection (“loss phase”) followed by a “plateau phase” when worm numbers remained stationary and a third “expulsion phase” when worm numbers again declined logarithmically with time. Irradiation of infective larvae at 50 and 100 kilorads (krad) affected the subsequent pattern of infection by prolonging the initial “loss phase” and contracting the “plateau phase”. The rate of decline of total worms during either the “loss phase” or the “expulsion phase” was not affected by larval irradiation.Few males were recovered at any stage of infection with larvae irradiated at 50 or 100 krad. However, the irradiated larvae (females) which survived continued development at a rate which was similar to that of normal larvae and it was apparent that these parasites were not able to maintain their position in the small intestine, but continued development during migration towards the posterior.The radiosensitivity of male larvae and worm egg production (eggs in utero) over a range of radiation doses (25 to 100 krad) were remarkably similar. On the other hand, female larvae were clearly less sensitive to γ-radiation. These observations suggested that the depressed egg production in females may be due to the absence of males.     

The effects of benzimidazole resistance and route of administration on the uptake of fenbendazole and thiabendazole by Haemonchus contortus and Trichostrongylus colubriformis in sheep

Radiolabelled fenbendazole (FBZ) and thiabendazole (TBZ) were administered into the rumen or abomassum of sheep previously infected with benzimidazole-susceptible or resistant Haemonchus contortus and Trichostrongylus colubriformis. Plasma radioactivity was determined for 12 hours following anthelmintic administration and at the end of this time the incorporation of radiolabel into the parasites was determined.Incorporation of radiolabel from FBZ was significantly greater in susceptible than in resistant worms. However, with TBZ there was no significant difference. There were no significant differences in the incorporation between H. contortus and T. colubriformis. Incorporation of TBZ was greater than FBZ in the worms except after ruminal administration to sheep showing clinical parasitism resulting from infection with benzimidazole- susceptible worms. TBZ was administered at five times the dose rate as FBZ, and thus a lower proportion of the administered TBZ than FBZ was incorporated.With the exception of incorporation of TBZ by resistant worms, the route of administration had a major effect on incorporation by the parasites and on plasma levels. Based on plasma levels, TBZ appeared to be rapidly absorbed from both the rumen and abomasum, whereas FBZ was less readily absorbed from the rumen with the rate of absorption being markedly greater if FBZ was administered directly into the abomasum. The findings suggest that the rumen may act as a reservoir of FBZ prolonging the period of high anthelmintic concentration in the host. This may contribute to the high efficacy of this anthelmintic.     

大鼠感染微小隐孢子虫后小肠、肾及肺脏病理学变化

将40只SPF级SD大鼠随机均分为地塞米松免疫抑制感染组和对照组n=20,免疫抑制感染组大鼠一次性接种1×106个/L微小隐孢子虫卵囊液1mL,空白对照组接种1mL灭菌生理盐水。于试验结束后宰杀大鼠,立即剖检取小肠、脾、肺,Bouin液固定,常规石蜡切片,HE染色,奥林巴斯显微摄影系统观察并显微摄影。结果显示,免疫抑制感染组大鼠小肠绒毛和肠腺受损严重,数量明显减少,结构模糊,绒毛上皮细胞和肠腺细胞明显肿胀、解体,细胞结构消失;肾脏肾小体数量减少,有的血管球明显萎缩甚至碎解,肾小囊腔明显增大,有的肾小囊壁层细胞的连续性遭到破坏;肺脏肺泡隔明显增厚,充血现象严重,肺泡隔内的细胞萎缩,胞核固缩,细胞结构模糊。试验结果表明,微小隐孢子虫对大鼠小肠、肾和肺等器官的组织结构具有明显损伤作用,进而影响大鼠的消化吸收、泌尿和呼吸等功能。     

Occurrence of 'attaching and effacing' lesions in the small intestine of calves experimentally infected with bovine isolates of verocytotoxic E coli

Nine calves (five colostrum-fed, four colostrum-deprived) were challenged with two field strains of Escherichia coli which produced either verocytotoxin 1 (VT1) or verocytotoxin 2 (VT2). Although three colostrum-fed calves had blood and mucus in their faeces, no diarrhoea was observed. Three of the four colostrum-deprived calves had diarrhoea and in two of them severe lesions were detected in the small intestine. Focal changes were detected in the colon of three calves. E coli were associated with the lesions in the small and large intestine and were shown by transmission electron microscopy to be intimately attached to the enterocyte surface with effacement of microvilli (attaching and effacing lesions). This is the first report of E coli which produce VT2 being associated with disease in calves.     

Interleukin-2 and interleukin-10 gene expression in calves experimentally infected with Fasciola gigantica

The gene expression of interleukin-2 and interleukin-10 in calves with a primary infection of Fasciola gigantica was studied. Five calves were infected orally with a dose of 1000 viable metacercariae of F. gigantica and five calves served as uninfected control animals. Expression of two cytokine genes i.e. IL-2 and IL-10 (Th1 and Th2) was measured at 10, 30 and 75 days post-infection (PI) by real-time polymerase chain reaction with the double stranded DNA-binding dye SYBR Green. Interleukin-2 was not detected in the peripheral blood mononuclear cells (PBMCs) of infected or control animals at 10, 30 and 75 days PI, however, IL-10 was present in detectable levels in PBMCs of infected animals at 10, 30 and 75 days PI, with no expression of this cytokine in the control animals. With an increased expression of IL-10 and no expression of IL-2 cytokine gene, the present study suggests that F. gigantica infection in calves evoked Th2 immune response.     

Myoelectric activity of the small intestine in enterotoxin-induced diarrhea of calves.

Electrodes were surgically implanted at 15-cm intervals in the jejunum and ileum of 4 healthy neonatal calves so that myoelectric activity could be recorded on 2 consecutive days. On the first day, each calf received a control treatment, and myoelectric activity was recorded for 340 minutes. Phase I was recorded for a mean of 175.8 +/- 22.8 minutes (51.5%), phase II for 124 +/- 27.4 minutes (36.5%), and phase III for 40.3 +/- 6 minutes (11.9%). On the second day, each calf was treated with approximately 200 micrograms of heat-stable enterotoxin (STa) of Escherichia coli orally. All calves developed diarrhea after the administration of STa. Phase I was recorded for a mean of 92.5 +/- 42.3 minutes (27.2%), phase II for 227.3 +/- 52.5 minutes (66.9%), and phase III for 20.3 +/- 11.4 minutes (6.0%). Increase in phase II and decrease in phases I and III after STa administration were significant (P less than 0.05). Duration of the migrating myoelectric complex was longer after STa administration (median, 64 minutes), compared with the control treatment (median, 54 minutes). Minute rhythms, recorded on the day of toxin administration, ranged from 49 to 153 minutes. There was no difference between the number of migrating action potential complexes on the control days (range, 1 to 10), compared with those on treatment days (range, 1 to 14). These findings are suggestive that enterotoxin-induced diarrhea of calves is accompanied by increased total spiking activity and minute rhythms in the distal portion of the jejunum and ileum.     

Histochemical distribution of enzymes in the small intestine of young milk-fed calves

The histochemical distribution of selected enzymes were examined in the small intestine of 5 about 3-week-old normal calves fed on whole cow’s milk. Alkaline phosphatase and β-D-galactosidase (= lactase) in the epithelial brush border, and non-specific esterase in the cytoplasm showed a strong reaction in the villi of the anterior small intestine and a marked decrease in the posterior regions. Aminopeptidase in the brush border of the villi showed a reverse distribution, with the strongest reaction in the posterior small intestine. Adenosine-triphosphate-splitting enzyme in the epithelial brush border, acid phosphatase and succinate dehydrogenase in the cytoplasm of the epithelial cells gave a relatively uniform reaction in the villi throughout the small intestine. A fluoride-resistant acid phosphatase was demonstrated in the brush border of the villi in the anterior small intestine. The distribution of enzymes demonstrated in this study was generally compatible with the known absorptive functions of the various parts of the small intestine.     

Demonstration of IgA-containing cells in the small intestine of lambs infected with Nematodirus battus

A peroxidase anti-peroxidase immunohistochemical technique was used to demonstrate IgA-containing cells in the small intestine of lambs infected with Nematodirus battus. These cells were more numerous in the infected lambs than in the uninfected animals. The difference was greatest for the first three sites, 1 to 3 m distal to the pylorus, where the preponderance of the N battus infection is found. It is suggested that future immunohistochemical studies on the relationships between IgA and resistance to N battus should be directed to this area.     

绵羊自然感染贝氏莫尼茨绦虫对小肠黏膜免疫相关细胞的数量变化

运用组织学和组织化学染色法,分别对自然感染贝氏莫尼茨绦虫的成年绵羊（感染组）与正常成年绵羊（对照组）的小肠各肠段的上皮内淋巴细胞、浆细胞、杯状细胞和嗜酸粒细胞的分布和数量变化进行了比较研究。结果显示：感染组小肠各段上皮内淋巴细胞、浆细胞、杯状细胞和嗜酸粒细胞数量均显著高于对照组,其中感染组十二指肠、空肠和回肠的上皮内淋巴细胞数量分别比对照组增加了169.11%、230.38%和233.42%（P〈0.01）;嗜酸粒细胞数量分别比对照组增加了116.78%、123.87%和164.51%（P〈0.01）;浆细胞数量分别比对照组增加了127.34%、72.97%和328.26%（P〈0.01）;杯状细胞数量分别比对照组增加了33.40%、41.42%和133.17%。对照组和感染组上皮内淋巴细胞和杯状细胞从十二指肠到回肠均逐渐减少,相反,对照组和感染组嗜酸粒细胞数量从十二指肠到回肠均逐渐增多,对照组小肠固有层浆细胞数量从十二指肠到空肠增加,空肠到回肠减少,感染组小肠固有层浆细胞数量从十二指肠到回肠均逐渐增多;对照组十二指肠、空肠、回肠的上皮内淋巴细胞、上皮内杯状细胞、浆细胞和嗜酸粒细胞均差异极显著（P〈0.01）;感染组十二指肠、空肠、回肠的上皮内杯状细胞、浆细胞和嗜酸粒细胞均差异极显著（P〈0.01）,感染组上皮内淋巴细胞空肠与回肠差异极显著（P〈0.01）,但十二指肠与空肠差异不显著（P〉0.05）。研究结果表明,贝氏莫尼茨绦虫感染成年绵羊后,成年绵羊通过特异性黏膜免疫细胞上皮内淋巴细胞增生加强细胞免疫水平,浆细胞增生加强体液免疫水平,同时还通过非特异性黏膜免疫细胞,嗜酸粒细胞和杯状细胞的增生进一步加强黏膜免疫水平以抵抗贝氏莫尼茨绦虫对绵羊的感染。可见绵羊可以通过黏膜免疫相关细胞增生加强局部免疫力以监视虫体免疫逃逸来抵抗寄生虫的感染。