Recently, it received much attention that the decline of fertility of eggs occurs in consequence of the application of artificial insemination during long period. It is not apparent what is the causative factor that might effect on fertility, but some investigators report that no fertilized eggs are due to antibody against spermatozoa, namely agglutinin.
This examination was undertaken to clarify a relation with such subjects.
The results obtained are as follows:
1. Through application of artificial insemination during long period a decline of fertility occurs exactly in some domestic hens, especially in heteroplasmic mating by White Cornish males distinctly in large number of cases.
2. Spermatozoa have antigenicity. By application of insemination agglutinin is produced without concerned to length of insemination period. Therefore, in hens with lower fertility and also with higher the agglutinin is similarly produced.
3. It is not able to be recognized that such a agglutinin has a significant effect to produce no fertilized eggs.

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1. The effect of different freezing point depressions (_??_) of saline solutions (estimated _??_: -0.76, -0.72, -0.68, -0.64, -0.60, -0.56, -0.52 and -0.48°C) on percentages of deformity and motile sperm were studied. Pooled semen from White Leghorn males was washed twice and resuspended to the original four volumes, and stored for 1 hour at 0-5°C. The deformity and number of motile sperm were reduced gradually as the saline solution became more hypertonic, but in the hypotonic solutions (_??_: -0.52 and -0.48°C) the deformity markedly increased.
2. The effect of different _??_ (-1.15, -0.91, -0.59 and-0.46°C) of glycine solutions on percentage of motile sperm was studied. Immediately after fresh semen was diluted 1 to 4, the percentage of motile sperm was 0 and 15% at _??_ of -1.15 and -0.91°C, respectively, but it increased to 30% at both _??_ after storage for 20 hours at 0-5°C. In contrast, the number of motile sperm was markedly reduced at _??_ of -0.59 and -0.46°C after storage for 20 hours.
3. Comparative studies of fresh and stored semen (24 hours at 0-5°C) on the response of spermatozoa to hypertonic glycine solution (_??_=-1.15°C) and hypertonic fructose solution (_??_=-1.28°C) were conducted at room temperature. The motility of spermatozoa was markedly but reversibly reduced in fresh semen diluted with the hypertonic glycine solution. Only a few spermatozoa were motile after storage for 4 hours, whereas the motility of spermatozoa diluted with the hypertonic fructose solution was markedly restored with the lapse of time and the percentage of motile sperm increased to 85% from the first 10% after storage for 1 hour.
The response of stored spermatozoa to the hypertonic glycine solution was much less than that of fresh spermatozoa.
4. An osmotic pressure difference between inside and outside of the sperm cell when semen was diluted 1 to 4 with a hypertonic glycine solution (_??_=-1.15°C) was estimated about 5, 6atm. at which most of the spermatozoa became reversibly immotile within a few seconds.

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Chromium (Cr) was often detected in the commercial fish meal and it was suspected that some foreign substances containing Cr was blended in the fish meal. In this paper, effect of feeding chicks the diets containing various types of Cr sources at different levels was studied.
In Expt. 1, four groups of 50 new-born White Leghorn chicks were fed experimental diets, respectively, supplemented 0, 100, 300 and 900p.p.m. of Cr as K₂CrO₄, which was used as a typical hexavalent Cr source. Feed intake and body weight gain for 6 weeks of chicks on 100p.p.m. Cr were almost equal to those of the control chicks on 0p.p.m. Cr. Feed intake and weight gain of chicks on 300p.p.m. Cr were 84.7 and 81.4% of those of the controls, respectively. Those of chicks on 900p.p.m. Cr were only 59.5 and 50.8% of those of the controls, respectively.
In Expt. 2, 4 groups of chicks similar to the previous experiment were fed 4 experimental diets supplemented 0, 300 and 900p.p.m. of Cr as chromium tanned hide powder, and 300p.p.m. of Cr as Cr₂(SO₄)₃. The former was the most suspected substance in fish meal and the latter was used as a typical trivalent Cr source. Feed intake and weight gain of chicks on hide powder was comparable to those of the controls regardless to the dietary Cr content. Feed intake and weight gain for 6 weeks of chicks on 300p.p.m. Cr of Cr₂(SO₄)₃ were 95.2 and 89.2% of those of the controls, respectively. The data suggested that poisonous effect of trivalent Cr was much less than that of hexavalent Cr.
Chromium content of various tissues and organs of chicks was determined with alkalifusion method modified slightly in the preparatory process. Chromium content in mucous membrane of gizzard of chicks on K₂CrO₄ was the highest and those in liver and kidney was higher than those of the control chicks. Accumulation of Cr in gizzard mucus membrane was also observed in chicks fed hide powder or Cr₂(SO₄)₃, although the amount of Cr was much less than that in chicks fed K₂CrO₄.

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The studies on the behavior of spermatozoa in the oviduct of the hen are important in order to clear the mechanism of retention for a long period of the fertilizing capacity of spermatozoa in the oviduct.
In the previous papers, the author stated that the inseminated spermatozoa were retained in the glandular cavities (sperm-nests) of both the infundibulum and the utero-vaginal junction, and the infundibulum probably had higher capacity for the preservation of spermatozoa than the utero-vaginal junction.
This paper describes the results concerning the fertility and spermatozoal behavior following infundibulal and vaginal inseminations, and the estimation concerning the difference in the capacity for the preservation of spermatozoa between the infundibulum and the utero-vaginal junction.
1) Inseminations of one hundred million spermatozoa were made in two sites; the infundibulum following laparotomy and the vagina. The weekly percentages of fertile eggs during three weeks following inseminations were respectively, 93.8, 88.2, 33.0 in the case of the infundibulum; 87.5, 50.0, 7.2 in the case of the vagina. The duration of fertility following the infundibulal insemination ranged over 12 to 27 days, with a mean of 17.2 with a S.D. of 4.0 days. On the other hand, when the insemination was made in the vagina, it ranged over 6 to 17 days, with a mean of 12.2 with a SD. of 3.5 days.
There appeared at 2nd week significant differences in percentages of fertile eggs following the infundibulal and vaginal inseminations, and so did they at 3rd week. And the durations of fertilities in the two cases of inseminations were different significantly.
2) Histological evidence showed that spermatozoa deposited in the infundibulum entered the infundibulal glands (sperm-nests) in all examples, but some spermatozoa migrated caudally and entered the utero-vaginal glands (sperm-nests) in several examples.
3) There were no significant difference in the percentages of fertile eggs and in the durations of fertilities following vaginal inseminations between one hundred million spermatozoa and larger numbers (four hundred million and one thousand million spermatozoa).
4) The results in this paper and the previous papers showed that most spermatozoa deposited in the infundibulum were preserved in the glands (sperm-nests) of this portion and most spermatozoa deposited in the vagina were preserved in the utero-vaginal glands (sperm-nests).
5) It seems likely that the significant differences in the percentages of fertile eggs and in the durations of fertilities between the infundibulal and the vaginal insemination were owing to the difference of the capacity for the preservation between the infundibulal and utero-vaginal glands (sperm-nests).

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Available energy of mutton fat and poultry grease was determined by the bioassay technique described in the first paper⁸⁾ of this series of studies. Available energies of fancy tallow and yellow grease imported from U.S.A. were also determined to compare the nutritive value among these animal fats. True digestibility of these fats was obtained from the dietary fat and total excreted fat corrected for excreted fat from fat free basal diet. The bioassay technique was based on the linear relationship between dietary energy level and the growth rate of chicks. The available energy of fats was tentatively shown by total digestible nutrients (TDN) in this paper.
Two preparations of mutton fat were used in this study and unexpectedly it was found that the available energies as well as true dige stibilities of these two preparations. having similar melting point of 43-45°C., were signfiicantly different (P<5%). One preparation had 144% of TDN and its digestibility was 61while the other had 203% of TDN and digestibility of 83 which was higher than digestibility of fancy tallow (P<1%).
Addition of equal volume of soybean oil or of a quarter volume of the methyl ester of mutton fatty acid to the mutton fat of lower nutritive value was not effective to improve the digestibility and nutritive value of a mutton fat. Emulsification of mutton fat also had little influence on nutritive value.
Poultry grease had high nutritive value of 207% of TDN and 97% of digestibility, which was almost comparable to the nutritive value of soybean oil. The findings agree with those reported by Cullen et al.²⁾
Available energies of fancy tallow and yellow grease were 174 and 145% of TDN, respectively. Digestibilities of these fats were 71 and 46%, respectively.

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Biological assay technique was applied for the determination of available energy of cassava meal, which was imported from Thailand for the first trial. Standard curve was obtained with the growth rate of chicks fed standard diets of various energy levels. Corn starch and yellow corn in the standard diet was replaced with cassava meal and levels of soybean oil and fish weal were adjusted to set the dietary energy and protein levels. With the growth rate of chicks on diet containing cassava meal, available energy of cassava meal was calculated with electronic computer. The available energy was shown as percentage of total digestible nutrients as in the previous 3 papers^{2, 3, 11)}.
In Expt. 411, it was shown that cassava meal fed at the level of 32.3% had growth retarding effect to day-old White Leghorn chicks, although no chick died during the experimental period of 4 weeks. In Expt. 503, cassava meal was soaked in water overnight or autoclaved at 120°C. for 1 hour. These treatments were quite effective to improve the growth rate of chicks. Diets containing 10 and 20% raw cassava meal were also fed in this experiment and it was revealed that chicks on 10% cassava diet grew almost satisfactorily, but the growth rate of chicks on 20% cassava diet was much slower than expected. To confirm the observation, diets containing 10 and 15% raw cassava meal was fed to 4-week-old chicks in Expt. 505. The growth rate of chicks on 10% cassava diet was normal but that on 15% cassava diet was not.
It was revealed in these experiments that cassava meal had some growth inhibiting factor which will be removed by heat-treatment or water-soaking. The growth-retarding effect was not detected when cassava meal was fed at 10% level. Prussic acid in cassava mael was discussed as the most possible growth-retarding factor. Available energy of cassava meal, when fed at 10% level or autoclaved or soaked in water was estimated. 70% in average. It is ecommended to use cassava meal at the level of less than 10% in poultry feed.
P.S. after the preparation of the manuscript, the authors have received the reprints of papers of Dr. Vogt et al, ^{20, 21, 22)} in which it was reported that the growth rate of broiler chicks on the diet containing 10% of cassava meal was normal. The content of prussic acid in their sample²²⁾ was much less than 36ppm in the sample tested in this paper.

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Objective of this experiment is to clarify the effects of nitrofurans on the growth of quails (Coturnix coturnix japonica).
Three kinds of nitrofurans, furazolidone (FZD), nitrofurazone (NFZ), and nihydrazone (NHZ) were added to a basal corn-soybean type ration at rate of each 30g per 1, 000 kg of the ration. Crude protein content of the ration was 19.8% in the ration I, 21.6% in the ration II, and 26.5% in the ration III. The former two rations were considered the low plane of protein contents for the growth of quails.
From results of experiments I, II, and III, it might be appeared that FZD had a growth-stimulating effect on quails, which were fed with low protein diet. On the other hand, neither NFZ nor NHZ had influences on the growth of quails. However, NFZ was most effective to decrease mortality due to some possibly bacterial disease, characterized of diarrhea, during the period of growth.
In experiments I and II, quails fed with low protein diet, either ration I or ration II, failed to renewal of feathers on the back. NHZ added to the ration II completely prevented the occurence of bare backs in experiment II.
Growth stimulating effect of FZD is discussed, especially from the view of protein sparing action.

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By means of immunoelectrophoresis with anti-WL serum, the serum components of domestic fowls used (White Leghorn, Barred Plymouth Rock, New Hamdshire, Rhode Island Red and White Cornish) were compared.
(1) The serum of WL was fractionated into ten or twelve fractions (Plate I-1).
(2) The serum samples obtained from the three varieties (WL, BP and NH) showed same immunoelectrophoretic pattern. On the other hand, the pattern of the varieties (SR and WC) was different from that of the former group of the variety (Plate I-3).
(3) The difference of the pattern between two above-mentioned groups was only observed in the α₁ region.
(4) Moreover, by means of specific staining method and method for absorbed antiserum, the protein showing the difference was determined to be lipoprotein.
(5) The precipitarion in the immunoreaction of α₁-lipoprotein in WL-group to the corresponding antibody was remarkably stronger than that in the SR-group.
(6) The following two hypothesis may be settled for the reason of the abovementioned grouping: (1) quantitative difference of α₁-lipoprotein in serum between the two groups; and (2) the difference of the similarity on the antigenic conostrnction of α₁-lipoprotein to that of WL serum used in the standard.

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