NOTE Internal Medicine

Effect of Supplementation of Dry Cat Food with D,L-Methionine and Ammonium

Chloride on Struvite Activity Product and Sediment in Urine

Masayuki FUNABA1), Takayo YAMATE1), Yukako NARUKAWA1), Ken GOTOH2), Tsunenori IRIKI1),
Yoshikazu HATANO2) and Matanobu ABE1)

1)
Laboratory of Nutrition, Azabu University School of Veterinary Medicine, 1–17–71 Fuchinobe, Sagamihara-shi 229–8501 and 2)R & D
Center, Nihon Nosan Kogyo K.K. 5246 Takura, Tsukuba-shi 300–2615, Japan

(Received 30 May 2000/Accepted 10 November 2000)

ABSTRACT. Feeding dry foods supplemented with urine acidifier (D,L-methionine (Met) or ammonium chloride) decreased urinary pH and
struvite activity product in clinically normal cats. As a result, the number of struvite crystals in urine was greatly reduced. Supplemen-
tation with 3% Met but not 1% Met caused decrease in the urinary concentration of sediment, which resulted from a reduction in the
HCl-soluble fraction. The concentration of HCl-insoluble sediment was not affected by supplementation with the urine acidifier.
KEY WORDS: feline, urinary sediment, urine acidifier.
J. Vet. Med. Sci. 63(3): 337–339, 2001

Struvite uroliths are frequently seen in cats fed dry foods, crystals and the amount of sediment. After immediately
when urinary pH exceeds 7.0. Therefore, supplementation measuring the pH of freshly voided urine, sulfuric acid was
of dry food with a urine acidifier such as D,L-methionine added to the urine collection bottle to prevent electrolytes
(Met) or ammonium chloride (NH4Cl) has been suggested to from crystallization and to prevent possible loss of ammo-
prevent struvite uroliths, although an excess of the acidifiers nia.
could cause harmful effects such as depressed food intake Because of the limited number of cats in experiment 1,
and metabolic acidosis [4], but urinary pH is not the sole experiment 2 was conducted to examine the effect of
determinant of the occurrence of struvite uroliths. It is another urinary acidifier, NH4Cl, in 6 adult cats (two male
known that organic compounds including proteins and gly- and four female; mean body weight with standard error, 3.8
cosaminoglycans could also regulate struvite crystallization ± 0.4 kg). They were used in a 3 week cross-over trial.
as a matrix [3]. Tamm-Horsfall protein, which is glycopro-
tein at 95–100 kDa, is assumed to be one of the candidates Table 1. Dietary ingredients and composition of basal diets
for the matrix [3], and a protein at 95–100 kDa was detected Experiment 1a) Experiment 2
mainly in urinary sediment fraction [1], so that it would be
necessary to pay attention to this fraction as a risk factor for Ingredients (%)
uroliths. The present study was conducted to examine the Corn gluten meal 15.0
Soybean meal 14.0
effect of urine acidifier (D, L-Met or NH4Cl) on the amount Fish meal 5.0
of sediment in the urine of clinically normal cats. Taurine 0.1
A total of 12 adult cats, which were clinically normal on Corn 42.0
the basis of physical examination, were used. All the cats Wheat flour 10.0
were cared for according to the principles outlined in the Beef tallow 6.5
NIH Guide for the Care and Use of Laboratory Animals [8]. Vitamins and mineralsb) 2.0
NaCl 0.8
The diet and water were available on an ad libitum basis
Ca(PO4)2 2.6
throughout the study. Nutrients found in all diets used in CaCO3 0.5
this study exceeded the requirements for the maintenance of Flavorc) 1.5
mature cats [9]. Food was provided daily at 4 PM. The cats Composition (% of dry matter)
were kept in a temperature-controlled room (24 ± 2°C) with Crude protein 31.2 26.9
artificial light provided from 6 AM to 6 PM daily. Ca 1.86 1.61
In experiment 1, the effect of supplemental Met was P 1.02 0.95
examined. Six adult cats (mean body weight with standard Mg 0.13 0.11
error, 4.1 ± 0.4 kg) were divided into 3 groups of 2 cats (one a) Commercial dry cat food containing corn, fish meal, meat meal,
male and one female), and fed experimental diets for 2 wheat flour, rice flour, beef tallow, flavor, vitamins, minerals, and
weeks. They were fed diets supplemented with 0%, 1% or taurine. b) One kilogram of the vitamin and mineral mixture contains
22,500 IU of vitamin A, 35 g of vitamin E, 2.2 g of vitamin B1, 2.3 g of
3% Met to commercial dry food (Table 1). For the last 3 vitamin B2, 1.6 g of vitamin B6, 8.5 mg of vitamin B12, 20 g of
days of the 2nd week of feeding, urine was collected every nicotinic acid, 5 g of panthothenic acid, 22 mg of biotin, 185 g of
24 hr at 4 PM. During this period, first-voided urine after 5 chlorine, 10 g of inositol, 450 mg of folic acid, 600 mg of Mn, 6.5 g of
AM was also obtained to measure urinary pH, the number of Fe, 33 mg of Co, 420 mg of Cu, 500 mg of I, and 500 mg of taurine.
c) Spray dried fish extract.
338 M. FUNABA ET AL.

They were fed diets with or without NH4Cl (1.5% of the group tended to be lower than that in the control group. The
diet) (Table 1). The NH4Cl was supplemented by decreas- urinary concentrations of Mg2+ and total P were not changed
ing corn content. For the last 7 days of each sampling by the additional Met, in contrast to an 80–90% increase in
period, urine was collected as described in experiment 1. the NH4+ concentration due to the Met supplementation.
The intake of food and water was recorded every day. The 3% Met supplementation tended to decrease the urinary
Urine samples during the sampling period from each cat in concentration of PO43-, suggesting increased P excretion as
experiments 1 and 2 were pooled and stored at –20°C until titratable acids. As a result, pSAP tended to increase in the
analysis. The amount of mineral in the diets and urine and 3% Met group. The increase in pSAP, i.e., decrease in SAP,
urinary total ammonia were measured as described previ- suggests solubilization of struvite crystals [5], and, in fact,
ously [7]. According to Buffington et al. [5], the solubility the number of struvite crystals remarkably decreased in the
of struvite crystals was determined on the basis of struvite 3% Met group.
activity product (SAP, [Mg2+] × [NH4+] × [PO43-]). In esti- The urinary sediment concentration tended to be lower in
mating the SAP, Mg and P were assumed to exist as ionic the 3% Met group than in the control group. There was also
forms and inorganic forms in urine, respectively. In addi- a tendency for the HCl-soluble fraction in the 3% Met group
tion, NH4+ and PO43- concentrations in urine were estimated to be lower than that in the control group, which suggested
from urinary pH and total concentrations of ammonia and dissolution of preformed struvite crystals caused by the sup-
inorganic P in urine as described previously [7]. For conve- plementation with 3% Met, but the urinary concentration of
nience, the SAP is expressed as pSAP, the negative loga- the HCl-insoluble fraction was not affected by dietary Met
rithm of SAP, because the SAP value is usually less than content. These results suggest that the reduction in the
10-8. Preformed crystals are more easily solubilized with amount of urinary sediment was mainly due to the reduction
decreasing SAP, i.e., increasing pSAP. Crystals in freshly in the HCl-soluble fraction.
voided urine were counted by light microscopy. The Similar results were obtained in experiment 2, where
amount of sediment in freshly voided urine was determined; NH4Cl was added to the diet as a urine acidifier. Urinary pH
the sediment after centrifugation (12,000 × g for 20 min at was significantly decreased by supplementation with 1.5%
4°C) was air-dried for 24 hr at 45°C, and the resultant sedi-
ment was weighed. The sediment was further fractionated
Table 2. Daily intake and urinary excretion of cats fed a diet supple-
by using 1 N HCl, and after extraction with 1 N HCl for 24
mented with D,L-methionine
hr at room temperature, the HCl-insoluble sediment was
measured as described above. The amount of the HCl-solu- Met: 0% 1% 3% SEM
ble fraction of the urine was determined by subtracting HCl- Food intake (g/d) 62.7 55.6 62.6 15.0
insoluble sediment from total sediment. Consistent with the Water intake (ml/d) 129.7 105.4 117.4 31.5
results of previous studies [3, 10], protein at 95–100 kDa Urinary pH 6.82 6.86 6.12 0.35
was detected by SDS-PAGE only from urinary sediment Urinary concentration
(data not shown). In addition, this protein was further Mg2+ (mM) 7.0 6.8 7.4 2.3
detected in the HCl-insoluble fraction of the sediment but NH4+ (mM) 61.3 110.8 116.2 20.5
Total P (mM) 50.0 50.4 36.4 11.4
not in the HCl-soluble fraction (data not shown), although it
PO43- (µM) 0.60 2.49 0.04 1.39
was expected in view of the extraction procedure. pSAP 9.71 9.46 10.61 0.46
Data in experiment 2 were analyzed by ANOVA, with the Struvite crystals (no./µl) 172.5 113.7 2.3 103.0
General Linear Model procedures [11]. The model included Total sediment (mg/ml) 2.22 2.28 1.43 0.58
diet, animal and period. HCl-soluble 1.83 2.02 1.00 0.64
All cats used in every experiment appeared to be healthy HCl-insoluble 0.38 0.23 0.43 0.09
and did not manifest any clinical abnormalities throughout
the study. Table 2 shows daily intakes and urinary excretion
in cats fed diet supplemented Met. Daily food and water Table 3. Urinary excretion of cats fed a diet supplemented with
intake was similar in all groups at the end of the 2nd week of NH4Cl
feeding, although cats in the 3% Met group exhibited a tran- NH4Cl: 0% 1.5% SEM P<
sient decrease in food intake within the first week (data not
Urinary pH 7.34 6.29 0.06 0.001
shown). Fau et al. [6] also reported that the addition of 2 to Urinary concentration
4% L-Met to cat food resulted in reduced food intake, Mg2+ (mM) 6.9 6.1 4.0 NS
although adaptation occurred after 10 d except when 4% L- NH4+ (mM) 241.6 468.9 23.9 0.001
Met was added. In our previous study [2], food intake and Total P (mM) 50.2 45.9 4.1 NS
body weight decreased but adaptation did not occur when PO43- (µM) 5.57 0.22 0.82 0.001
6% Met was added to the diet. The transient decrease in pSAP 8.43 9.65 1.24 0.001
food intake resulting from an addition of 2 to 3% Met may Struvite crystals (no./µl) 447.9 80.0 24.1 0.001
be due to a Met imbalance rather than Met toxicity, which Urinary sediment (mg/ml)
HCl-insoluble 1.38 1.13 0.18 NS
could occur when more than 4% Met was added to the diet.
Urinary pH in the 3% Met group but not in the 1% Met NS: P>0.10.
 SEDIMENT IN FELINE URINE 339

NH4Cl, leading to an increase in pSAP mainly resulting Res. 55: 965–971.

from a decrease in the PO43- concentration (Table 3). Stru- 4. Buffington, C.A., Cook, N.E., Rogers, Q.R. and Morris, J.G.
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Cambridge, UK.
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