Genetic resistance to Ascaridia galli infections in chickens.

Ascaridiosis is still a cause of economic losses in modern poultry production systems. Improving genetic resistance to this disease provides an attractive alternative for free-range organic poultry producers and will eventually reduce the extensive use of anthelmintic drugs. An experiment was conducted to compare resistance to Ascaridia galli infections in Lohman Brown (LB) and Danish Landrace (DL) chickens. A self-cure mechanism to A. galli infections was observed in both breeds. However, significantly higher worm burdens and egg excretion were seen in the DL compared to LB chickens during primary infection. This indicates that breeding for resistance to A. galli is possible in chickens.     

Ascaridia galli infections in free-range layers fed on diets with different protein contents

1. One hundred and twenty 17-week-old Lohman Brown hens were divided into 4 groups. Groups 1 and 3 were given a diet with 180 g protein/kg and groups 2 and 4 were given a diet with 140 g protein/kg. Groups 1 and 2 were orally infected with 500 (50) embryonated Ascaridia galli eggs. 2. Marked differences in mean weekly weight gain for the 4 groups were observed. 3. Hens given 140 g protein/kg had a significant lower mean worm burden of adult A. galli worms and a significant lower weight gain compared to the group given 18 g protein/kg. 4. There was no significant difference in faecal egg counts between the 2 parasitised groups. 5. The egg production did not differ significantly between any of the groups. 6. The results of this study indicate that the amount of dietary protein in the diet has an effect on the establishment of A. galli infections in the gut of layers kept under free range conditions.     

Effect and repeatability of Ascaridia galli egg output in cockerels following a single low dose infection

Three groups of caged 20 Tetra-SL cockerels aged 1 day were orally infected with 30, 60 or 125 embryonated Ascaridia galli eggs. After 11, 12 and 13 weeks, faecal egg counts (FECs) were determined. All birds were slaughtered after the last sampling. A group of 25 control birds was sampled and slaughtered in parallel. The gastrointestinal tracts were examined for the presence of adult stages of A. galli. A random sample of 10% was also examined for the presence of immature stages of A. galli. The group with an infection dose of 125 eggs showed the highest average worm burden (p<0.05) and number of females (p<0.05), but the mean establishment rate was the lowest in this group. There was no significant difference in the mean logFEC between the groups. The logFEC per female worm was the highest in the low infection group (p<0.05). The average worm length and weight and the birds body weight were not significantly different among the groups. The estimated repeatabilities for mean logFEC of the different samples were reasonably high (0.55-0.87). This may open a way of genetic selection for A. galli resistance in chickens, which will be of importance for birds kept in alternative and organic farming systems.     

Age-related differences of Ascaridia galli egg output and worm burden in chickens following a single dose infection

Ninety white chickens (Lohmann LSL) were reared under helminth-free conditions and divided into five groups. Four groups were artificially infected with 250 embryonated Ascaridia galli eggs at the age of 6, 12, 18 or 24 weeks. Ten birds were kept as uninfected controls. Six and 10 weeks after infection (p.i.), individual faecal egg counts (FEC) were performed. The birds were slaughtered after the second sampling and their gastrointestinal tracts were examined for the presence of adult A. galli. The FEC increased from the first to the second sampling significantly in all the infected groups. The highest increase was shown in the group infected at 12 weeks of age, whereas the increase in the other groups was relatively moderate. However, the total worm burden and mean FEC at the second sampling were highest (p<0.01) in those birds infected at an age of 12 or 18 weeks. The serum protein and triiodothyronine (T3) levels did not differ significantly (p>0.05) between any of the groups. Thyroxine (T4) was significantly different between the groups infected at 6 and 18 weeks of age (p<0.05), and those at 6 and 24 weeks of age (p<0.01). The thyroid hormone levels correlated significantly with the FEC. Age does not seem to play a major role in resistance to A. galli infections in layers, whereas a bird's hormonal and immune status, related to laying activity, seems to have a significant negative impact on resistance.     

Comparative genetic resistance to Ascaridia galli infections of 4 different commercial layer-lines

1. The objective of the study was to compare the establishment and effect of Asaridia galli infections in 4 different layer-lines. 2. A total of 160 birds comprising 4 different commercial layer-lines, ISA Brown, New Hampshire, Skalborg and a cross of New Hampshire(NH) and Skalborg (Sk), were infected with A. galli eggs. The birds were examined for the presence of parasite eggs and parasites at weeks 3, 6 and 9 post infection (pi). 3. At week 6 pi the chickens of the NH line harboured more larvae compared with the three otherlines. The Sk line chickens excreted more A. galli eggs throughout the study compared with the other lines. Female worms in the Sk line were more fecund than the worms in the other lines. Male and female worms recovered from the Sk line at week 9 pi were longer. Male worms recovered from the NH line 6 weeks pi were shorter than male worms from the other lines. Female worms recovered from the NH line were shorter than the female worms from the ISA line and the Sk line. No differences were seen in weight gain among the 4 lines. 4. The results suggest that genetic factors are involved in the establishment and survival of A. galli in the intestine of layers. Further studies are needed to elucidate the genetic mechanisms behind the observed parasitological findings.     

The effect of concurrent infections with Pasteurella multocida and Ascaridia galli on free range chickens

Pasteurella multocida and Ascaridia galli are observed with high prevalences in free range chickens in Denmark, but the impact is unknown. A study was carried out to examine the interaction between A. galli and P. multocida in chickens and the impact on production.Five groups, each with 20 18-week-old Lohmann Brown chickens were infected. Group 1 was orally infected with 1000+/-50 embryonated A. galli eggs. Group 2 received 10(4) cfu P. multocida intratracheally. Group 3 was infected with A. galli and subsequently with P. multocida. Group 4 was infected with P. multocida followed by A. galli. Group 5 was the control. The study ran for 11 weeks where clinical manifestations, weight gain and egg production were recorded. Excretion of P. multocida was determined on individual basis and blood smears were made for differential counts. At the end of the study pathological lesions and the number of adult worms, larvae and eggs in the faeces were recorded.The birds were more severely affected when infected with both pathogens compared to single infections with A. galli or P. multocida, respectively. A lower weight gain and egg production was observed with dual infections. A. galli infection followed by a secondary P. multocida infection resulted in more birds with pathological lesions and continued P. multocida excretion.In conclusion a negative interaction between A. galli and P. multocida was observed and it is postulated that free range chickens are at higher risk of being subjected to outbreaks of fowl cholera when they are infected with A. galli.     

Anthelmintic activity of ivermectin against experimental Ascaridia galli infection in chickens

The efficacy of ivermectin against Ascaridia galli infection was evaluated in chickens under controlled laboratory conditions in two separate experiments. In each experiment 22 White Leghorn chicks were randomly assigned to three groups of 10 (infected-treated), 9 (infected-control) and 3 (uninfected-control) birds. Each bird in the former two groups was orally infected with 1,500 embryonated A. galli eggs. The chicks in the treated group were subcutaneously injected with ivermectin at a dose of 0.3 mg kg-1 body weight on Day 10 (Experiment 1) and Day 35 post-infection (Experiment 2) for immature and adult infections, respectively. The treated birds had 0.9% (Experiment 1) and 0.4% (Experiment 2) worm recovery compared with 8.7 and 8% in the infected-untreated controls of the respective experiments. The fall in post-treatment faecal egg counts was 81 and 92% in birds treated on Days 10 and 35, respectively. The drug was found to be 90 and 95% effective against immature and adult worms, respectively. The lower lesion score and post-treatment near-normal haematobiochemical picture in treated birds confirmed these observations. The treated birds also had a better growth rate than the untreated chickens. The mature worms in the intestinal lumen of the host were more sensitive to the treatment than the immature stages of the parasite in the tissue phase.     

Establishment of Ascaridia galli in betamethasone-treated chickens

Twenty-five day-old White Leghorn chickens were each infected orally with 500 (Group I), 1000 (Group II) and 2000 (Group III) infective eggs of Ascaridia galli and were killed 30 days after the infection. A high percentage of the infecting dose (6.5%) established as adult worms in the intestine of chickens receiving the lowest level of primary infection, but as the amount of primary infection given to birds increased, there was a significant fall in the percentage establishment of adult worms in the intestine. A similar pattern of worm establishment was shown by chickens of the same age and receiving similar levels of primary infections, but which were treated with betamethasone at a dose of 2 mg/kg body weight commencing 5 days before and continuing up to 15 days after the infection. Betamethasone-treated birds, however, showed more establishment of worms in the intestine, but lower weight gains in comparison to the birds which were not treated. Different levels of primary infections given, with or without treatment with betamethasone, had no effect on the sex ratio of the resultant male/female worm populations, which became established in almost equal numbers in the intestine of chickens.     

Tetrameres americana Cram (1927) populations in chickens infected with different dose levels

Three groups of 20-45 Lohman brown chickens aged 3 weeks were orally infected with doses of 25, 100 and 400 Tetrameres americana, respectively. Fifteen chickens were kept as uninfected controls. Every third week in a period of 12 weeks, 5-15 chickens were slaughtered and the proventriculi were examined for the presence of adult stages of T. americana. From day 21 post-infection, pooled feacal samples were examined for parasite eggs, whereas the weight gain of the chickens was monitored weekly. The parasite established the infection in similar rates in all the three groups, 9.5-15.2%, except on day 84 post-infection, when the establishment rate of the high-dose group was significantly lower (P < 0.005). The average worm burden increased with increasing dosages, though displaying the worm burden of the high-dose group as being roughly halved on day 84 post-infection, thus suggesting an expulsion of worms. Females were more abundant than males. The mean lengths of male and female specimens showed no significant differences between the groups. The egg output was also increased with increasing dosages with the earliest prepatent period of 38 days post-infection found in the high-dose group. Infected chickens exhibited no difference in weight gain in comparison with the controls. This study demonstrated that single infections with varying doses influenced the establishment rate and the worm burden but not the parasites egg excretion, worm size nor the weight gain and mortality of the final host.     

Influence of Ascaridia galli infections and anthelmintic treatments on the behaviour and social ranks of laying hens (Gallus gallus domesticus)

In the present study, the effects of an experimental Ascaridia galli infection and anthelmintic treatment on the behaviour and social status of laying hens of two different lines were studied. Sixty white (Lohmann LSL; LSL) and 60 brown (Lohmann Brown; LB) hens were reared under helminth-free conditions. The hens of each line were divided into four groups. The birds in two of the groups were artificially infected with 250 embryonated A. galli eggs at an age of 27 weeks. The other two groups were kept as uninfected controls. One infection and control group was dewormed at 38 weeks of age and slaughtered 4 weeks later, contemporary with the other animals. Individual faecal Ascaridia egg counts (FEC) were performed 11 weeks post-infection (p.i.). Body weights, laying performance and egg weights were recorded regularly. Blood was taken to measure testosterone levels. The worm burdens established in the intestines were counted in the infected not treated group after slaughtering. In addition, 15 behavioural parameters were recorded by focal animal observation (n=10 per group) of one infection (plus anthelmintic treatment) and one control group, according to the time-sampling method throughout the experiment. All agonistic interactions within the groups were recorded simultaneously on an ongoing basis, thereby allowing the calculation of an individual social rank index. The following results were obtained: Mean FEC and worm burden were higher (p < 0.01) in the LSL hens than in the LB hens, but their performances were not different (p > 0.05) from the controls. Infections with A. galli resulted in significant behavioural changes in both lines as the infected birds showed a higher food intake and lower locomotion activity during the prepatent and patent periods. After anthelmintic treatment, food intake decreased and locomotion increased. Behavioural changes were more pervasive in the infected LSL hens, as these hens also showed changes in ground pecking and nesting activity not only during the prepatent and patent periods, but also after anthelmintic treatment. Social rank did not significantly change as a consequence of A. galli infection, but the infection groups of both lines had a tendency to display more agonistic activity than the non-infected controls. Serum testosterone values only increased significantly in the LSL hens during the patent period compared to the controls. No significant correlations were apparent between serum testosterone level, parasitic parameters, social status and/or aggressiveness in any of the groups. This study showed that even sub-clinical A. galli infections and their anthelmintic treatment can have an impact on animal behaviour.     

The effect of Plasmodium gallinaceum on a challenge infection with Ascaridia galli in chickens

The effect of a primary infection with the haemoparasite Plasmodium gallinaceum on the establishment of a challenge infection with the nematode Ascaridia galli in chickens was studied. Four groups were infected as follows. Group 1: inoculated intravenously with 10(6) P. gallinaceum-infected erythrocytes on day 0; group 2: orally infected with 500 embryonated A. galli eggs on day 10; group 3: infected with P. gallinaceum on day 0 and A. galli on day 10; and group 3: non-infected control birds. The results of this investigation demonstrates that a primary infection with P. gallinaceum in chickens alters the course of a subsequent infection with A. galli. Thus, an antagonistic effect was seen in which the malaria infection caused a significant reduction on the establishment of the nematode in concurrently infected animals.     

Genetic resistance to Heterakis gallinarum in two chicken layer lines following a single dose infection

Groups of White Leghorn and New Hampshire male and female chicks were reared under helminth-free conditions and divided into two groups at an age of 8 weeks. One group was artificially infected with 100 embryonated Heterakis gallinarum eggs, while the other group was kept as uninfected control. After 8 weeks all animals were harvested and worm burden was established. Maximum of 60 worms per bird randomly selected were measured for length. Body weights were recorded at 8 and 16 weeks of age. Total worm burden was significantly (P<0.0001) higher in infected males than in infected females. Furthermore, White Leghorns showed a significantly (P=0.011) higher number of worms when compared with New Hampshire animals. Average worm lengths were not significantly different between the breeds (P=0.155) and sexes (P=0.062). Correlations between worm burden and worm length was 0.08 (P=0.140). The daily weight gains between 8 and 16 weeks of age of the infected animals were significantly (P=0.038) lower when compared with the control group. There was no significant correlation between daily weight gain and worm burden. The estimated heritabilities for logarithm (ln) worm burden were 0.41 (S.E.+/-0.086) in White Leghorn and 0.31 (S.E.+/-0.126) in New Hampshire, respectively. Based on the estimated heritabilities it is possible to select for H. gallinarum resistance in both genetically distinct chicken breeds. This approach may be of importance for chickens kept in floor husbandry systems.     

数种驱虫药和化学药物对体内外鸡蛔虫产卵和虫卵发育的影响

为鉴定男性不育药和有些驱虫药对寄生蠕虫的不育作用,对培养在试管内的鸡蛔虫和卵胚,以及人工感染蛔虫的鸡分别进行了体内和体外试验。棉酚和硝基呋喃类的呋喃唑酮、呋喃西林、S_(73031)和S_(72055)均无抑制产卵和卵胚发育的作用,苯并咪唑类的丙硫咪唑、甲苯咪唑和噻苯咪唑以及吩噻嗪均有抑制产卵和卵胚发育的作用,它们的EMIC_(50)分别为0.006,1.326,0.029和0.052ppm。抑制产卵所需浓度远大于抑制卵胚发育所需浓度。吡喹酮和硫双二氯酚也不具有抑制产卵和卵胚成熟的作用。放线菌酮仅对试管内蛔虫有抑制产卵作用,但对体内蛔虫却无影响。     

Anthelmintic effects of citrus peels ethanolic extracts against Ascaridia galli

The use of phytogenic bioactive compounds to control poultry helminthes is increasing in different production systems. In vitro and in vivo anthelmintic activity of citrus peels against Ascaridia galli was investigated. Ethanolic extracts of three citrus peels species were suspended in 0.5% dimethyl sulfoxide (DMSO) to form an experimental composition (EC). EC was mainly composed of Limonene (96%), followed by β-Pinene (1.5%), α-Pinene (0.5%), and Sabinene (0.3%). For in vitro investigation, adult A. galli worms (n=225) were collected from naturally infected chickens and distributed to 3 equal groups. Groups 1, 2, and 3 were exposed to Fenbendazole (0.5mg/ml), EC (50mg/ml), and 0.5% DMSO, respectively. For in vivo investigation, 200 Lohmann Selected Leghorns chicks were infected at 1-day old with 250 embryonated A. galli eggs. At 6 weeks of age, 150 A. galli infected birds were randomly allocated into 5 equal groups. Groups 1, 2, and 3 were treated with 300, 600, and 1200 mg EC kg(-1) body weight, respectively. Group 4 was treated with Fenbendazole (50 mg kg(-1)). Group 5 was left as control. Birds were euthanized 2-weeks post-treatment, and all worms were collected from their intestines. EC possessed significant (P<0.001) in vitro anthelmintic properties on live worms. No significant (P>0.05) difference was quantified between number of motile worms exposed either to EC or Fenbendazole 7h post-exposure. A significant (P<0.0001) reduction in fecal egg count was observed 14 days post-treatment with 1200 mg kg(-1) EC. No significant differences were observed in worm burden of the 300 mg EC-treated group compared to the controls. In contrast, the 600 and 1200 mg EC-treated groups showed significant (P<0.0001) reduction in worm burden. Fenbendazole was the most effective in reducing A. galli burden (Efficacy=97%) followed by 1200 mg EC kg(-1) (68%), 600 mg EC kg(-1) (66%), and 300 mg EC kg(-1) (5%). It is concluded that citrus peels extracts have potential anthelmintic properties against A. galli.     

Genetic differences of Ascaridia galli egg output in laying hens following a single dose infection.

Groups of 20-week-old white (Lohmann LSL) and brown (Lohmann Brown) hens were reared under helminth-free conditions and both divided into two subgroups. One subgroup was artificially infected with 250 embryonated Ascaridia galli eggs, the other subgroup were kept as uninfected controls. During the following 12-month laying period individual faecal Ascaridia egg counts (FEC) were performed and bodyweight measured at monthly intervals. Laying performance and egg weights were determined daily. The mean FEC were significantly (P < 0.01) higher in infected white hens than in infected brown hens. The growth rate of the white hens was significantly (P < 0.05) higher in the control group, whereas in brown hens no significant group difference was observed. The laying performances and egg weights did not differ significantly (P > 0.05) between control and infected animals for either line. The estimated repeatabilities for mean log(10) FEC of different samples were reasonably high (0.16-0.54). Heritabilities for mean log(10) FEC were between 0.13 and 0.19 for white hens and between 0.0 and 0.10 for brown hens. Thus, it should be possible to select for A. galli resistance in chickens, which will be of importance for birds kept in alternative and organic farming systems.     

Duration of immunity induced in chickens by an attenuated live Salmonella enteritidis vaccine and an inactivated Salmonella enteritidis/typhimurium vaccine

The aim of this study was to examine the duration of immunity of different vaccination schemes using the S. enteritidis live vaccine Gallivac Se and the S. enteritidis-S. typhimurium inactivated vaccine Gallimune Se+St. Three groups of Lohman Brown chickens were used. Group one was vaccinated three times orally with Gallivac Se at weeks one, seven and 13 of age. Group two was vaccinated twice orally with Gallivac Se in weeks one and seven and once i.m. with Gallimune Se+St in week 14 of age. A third group was not vaccinated and served as the control group. Eight randomly selected chickens from each of the three groups were challenged with a nalidixic acid resistant S. enteritidis PT4 strain in weeks 24, 51 and 71 of age and the same number of animals were challenged with a S. typhimurium DT 104 strain in weeks 26, 54 and 73 (75) of age.The chickens were euthanised seven days post challenge and the number of challenge strain organisms (log10 cfu) in the liver and on caecal mucosa was determined.The quantitative investigation of the challenge strain in the liver and caecal mucosa revealed a statistically significant (p < 0.05) lower challenge strain burden in the vaccinated groups compared with the non-vaccinated control group up to week 71 (73) of age. The protective effects were demonstrated for both challenge strains.     

Genetic resistance to natural helminth infections in two chicken layer lines

Groups of Lohmann Brown (LB) and Lohmann Selected Leghorn (LSL) hens were reared under helminth-free conditions and kept afterwards together in a free range system. Mortality rate, body weight development, laying performance and faecal egg counts (FEC) were recorded during a 12 month laying period. At the end of the laying period, 246 LSL and 197 LB hens were necropsied and worms counted following the World Association for the Advancement of Veterinary Parasitology (W.A.A.V.P.) guidelines. In addition adult Heterakis gallinarum and Ascaridia galli were sexed and measured for length. Significant (P<0.01) differences were observed in mortality rates between LSL and LB animals (12.9 vs. 5.7%). LSL hens showed significantly (P<0.05) higher FEC when compared with LB hens at almost all dates of monitoring. Almost all animals became infected with at least one helminth species. The most prevalent species were H. gallinarum, Capillaria spp. and A. galli. LB hens showed a significantly (P<0.05) higher average number of adult H. gallinarum, Capillaria spp. and tapeworms when compared with LSL animals. However, number of adult A. galli was in tendency lower in these animals. In total, LB had a significantly (P<0.05) higher worm burden than LSL (192.3 vs. 94.3). The estimated heritabilities for total worm burden were 0.23 (SE±0.12) in LSL and 0.75 (SE±0.21) in LB, respectively. The number of all different helminth species were positively correlated. The sex ratio of H. gallinarum and A. galli and the average worm lengths were not significantly (P>0.05) different between the genotypes. There was no significant phenotypic correlation between body weight and worm burden in LSL, whereas it was the case in LB (r=0.17, P<0.05). Based on the estimated heritabilities it is possible to select for helminth resistance in both genotypes.     

Avian leukosis virus (ALV) infection, shedding, and tumors in maternal ALV antibody-positive and -negative chickens exposed to virus at hatching

Chickens highly susceptible to avian leukosis virus (ALV) infection and tumors, with and without ALV subgroup A maternal antibody (MAB), were infected with a field strain of ALV subgroup A at hatching. Viremia, antibody development, cloacal and albumen shedding, and tumors in chickens with MAB (MAB+) were compared with those in chickens lacking MAB (MAB-). At 18 weeks of age, the incidence of viremia was significantly lower in MAB+ chickens than in MAB- chickens; further, MAB significantly reduced the proportion of tolerantly infected (viremic antibody-negative) chickens. Cloacal shedding of ALV at 22 weeks of age and shedding of ALV group-specific (gs) antigen in albumen of eggs from all laying hens at 30-32 weeks of age were significantly lower in MAB+ hens than in MAB- hens. The incidence of ALV-induced tumors was lower in MAB+ chickens than in MAB- chickens, significantly so in one of three trials conducted. These results suggest that MAB may influence the development of viremia, antibody, and shedding of ALV following massive exposure to virus at hatching.     

饲养方式对卢氏绿壳蛋鸡屠体性状的影响

本研究选择笼养和放养条件下49周龄的卢氏绿壳蛋鸡各20只（公母各半）,测定了其屠体性状。结果表明,饲养方式对卢氏绿壳蛋鸡的活重、屠宰率、半净膛率、全净膛率、胸肌率、腿肌率、腹脂率、翅重、心脏重、肝脏重、脾重、肠重、胃重和生殖器官重的影响均差异不显著（P>0.05）。饲养方式对公鸡的各项指标影响差异不显著（P>0.05）,而对母鸡的头重、颈重、脚重影响很大（P<0.01）;相同饲养条件下公鸡的全净膛率、腹脂率、头重、脚重、颈重、翅重、心脏重和肠重极显著高于母鸡（P<0.01）,对肝脏重、脾脏重和胃重的影响差异不显著（P>0.05）;笼养时公鸡的活重、腿肌率和生殖器官重显著高于母鸡（P<0.05）,而对半净膛率和胸肌率的影响不显著（P>0.05）;放养时母鸡的生殖器官重极显著的高于母鸡（P<0.01）;不同饲养条件下,公鸡头重、颈重、肠重、生殖器官重与屠宰率相关性不显著（P>0.05）;母鸡脚重、翅重、肠重、脾脏重与屠宰率的相关性显著（P<0.05）。     

Failure of young goats to acquire resistance to Haemonchus contortus

Two experiments were carried out to investigate the acquisition by goats of resistance to Haemonchus contortus. In Experiment 1, five Saanen wethers reared worm-free and averaging 5 1/2 months of age at the start of the experiment, were dosed with 200 H. contortus infective larvae three times per week for 10 weeks (approximately 23 infective larvae/kg mean initial liveweight/week) and then given anthelmintic treatment. Each goat and an equal number of worm-free controls were then challenged with 10,000 infective larvae. Post mortem worm counts were carried out 30 days later. In Experiment 2, eight worm-free Saanen wethers, 14 months old at the start of the experiment, were dosed with 825 infective larvae per week for 14 weeks (approximately 23 infective larvae/kg mean initial liveweight/week) except for one week when only 300 larvae were given and one week when no larvae were given. After anthelmintic treatment, each received, together with seven worm-free control animals, a challenge dose of 15,000 infective larvae. Post-mortem worm counts were carried out 28 days later. There were no significant differences in post-mortem worm counts between previously infected and uninfected groups in either experiment. In both experiments, serum pepsinogen values rose significantly as a result of infection but there was no significant (p>0.5) correlation between worm counts and pepsinogen values on the day of slaughter.